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1.
Ciênc. rural (Online) ; 52(2): e20200894, 2022. ilus, tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1339655

ABSTRACT

Seed germination is a complex process controlled by many factors, in which physical and biochemical mechanisms are involved and the mobilization of reserves is crucial for this process to occur. Although, seed reserve mobilization is usually thought to be a post-germination process, seed reserve proteins mobilization occurs during germination. This study quantified seed proteins of bean genotypes during different hydration times, in order to understand the process of protein mobilization and whether there is relationship of this biochemical component with seed vigor. This study was conducted using seeds with different levels of vigor, genotypes with highest (13, 42, 55 and 81) and lowest (07, 23, 44, 50, IPR-88-Uirapurú and Iapar 81) physiological quality. High vigor genotypes showed greater efficiency in hydrolysis and mobilization of protein component, because they presented low globulins content in cotyledons at radicle protrusion in relation to low vigor genotypes (07, 23 and 50). The protein alpha-amylase inhibitor, observed in all genotypes, is involved with the longer time needed for radicle protrusion, according to the band intensity difference in genotypes 07, 44 and Iapar 81.


A germinação de sementes é um processo complexo controlado por muitos fatores, nos quais mecanismos físicos e bioquímicos estão envolvidos e a mobilização de reservas é decisiva para que esse processo ocorra. Embora a mobilização de reservas de sementes seja considerada um processo pós-germinativo, a mobilização das proteínas de reserva de sementes ocorre durante a germinação. Este estudo teve como objetivo quantificar as proteínas de sementes de genótipos de feijão durante os diferentes tempos de hidratação, a fim de compreender o processo de mobilização proteica e se há relação desse componente bioquímico com o vigor das sementes. Este estudo foi realizado utilizando sementes com diferentes níveis de vigor, genótipos com maior (13, 42, 55 e 81) e menor (07, 23, 44, 50, IPR-88-Uirapurú e Iapar 81) qualidade fisiológica. Os genótipos de alto vigor apresentaram maior eficiência na hidrólise e mobilização do componente proteico, pois apresentaram baixo teor de globulinas nos cotilédones na protrusão radicular em relação aos genótipos de baixo vigor (07, 23 e 50). A proteína inibidora da alfa-amilase, observada em todos os genótipos, está envolvida com o maior tempo necessário para a protrusão da radícula, de acordo com a diferença de intensidade da banda nos genótipos 07, 44 e Iapar 81.


Subject(s)
Seeds/chemistry , Genetic Variation/genetics , Proteins/analysis , Phaseolus/embryology , Mass Spectrometry , Electrophoresis, Polyacrylamide Gel
2.
Arch. latinoam. nutr ; 71(4): 241-251, dic. 2021. tab
Article in Spanish | LILACS, LIVECS | ID: biblio-1355032

ABSTRACT

El basul es el fruto de la planta Erythrina edulis que posee un alto contenido de proteínas, fibra dietética y antioxidantes, pero también con contenido de antinutrientes. Objetivo. El objetivo del estudio fue evaluar el efecto del germinado sobre las características nutricionales, propiedades bioactivas y funcionales de las semillas de basul. Materiales y métodos. Mediante un diseño aleatorizado con arreglo factorial de 2x2x2 (tiempo de remojo, tiempo de germinado, presencia o ausencia de luz), las semillas de basul fueron germinadas y convertidas en harina (HBG) para determinar la composición proximal y la digestibilidad in vitro de la proteína. Además, se ha determinado los fenoles totales (CFT), la capacidad antioxidante (métodos ABTS y DPPH) y las propiedades de hidratación y adsorción de aceite. Resultados. Las diferentes condiciones de germinado no han modificado la composición proximal de la HBG; sin embargo, la digestibilidad in vitro de la proteína incrementó hasta en 6,25% en uno de los tratamientos respecto a la muestra no germinada. El CFT, también ha incrementado de 241,49 mg AGE/100g (sin germinar) a 267,15 mg AGE/100g (germinado); al igual que la capacidad antioxidante (ABTS) de 173,04 µmolTE/g (sin germinar) a 195,67 µmolTE/g (germinado). También se tuvo incrementos en la solubilidad, la capacidad de absorción de agua y la capacidad de hinchamiento. Conclusión. La calidad de la proteína, el contenido de fenoles totales, la capacidad antioxidante y propiedades funcionales de interacción con el agua de la semilla de basul mejora con el germinado(AU)


Basul is the fruit of the Erythrina edulis plant. It has a high content of proteins, dietary fiber, and antioxidants, but it also contains antinutrients. Objective. The study objective was to evaluate the effect of sprouting on the nutritional characteristics, bioactive and functional properties of basul seeds. Materials and methods. Basul seeds were germinated to produce flour (HBG) and determine its proximal composition and in vitro digestibility of the protein through a randomized design with a 2x2x2 factorial arrangement (soaking time, germination time, and presence or absence of light). Total phenols (CFT), antioxidant capacity (ABTS and DPPH methods), and hydration and oil adsorption properties were also determined. Results. The different germination conditions did not modify the proximal composition of HBG. However, the in vitro digestibility of the protein increased up to 6.25% in one of the treatments compared to the non-germinated sample. The CFT also increased from 241.49 mg AGE/100g (without germination) to 267.15 mg AGE/100g (germinated) and the antioxidant capacity (ABTS) from 173.04 µmolTE/g (without germination) to 195.67 µmolTE/g (germinated). Solubility, water absorption capacity, and swelling capacity also increased. Conclusion. The quality of the protein, the content of total phenols, the antioxidant capacity, and the functional properties of interaction with the water of the basul seed improved with germination(AU)


Subject(s)
In Vitro Techniques , Dietary Fiber , Dietary Proteins , Germination , Erythrina , Phenolic Compounds , Flour/analysis , Proteins , Absorption , Fabaceae , Antioxidants , Nutritive Value
3.
Infectio ; 25(4): 270-275, oct.-dic. 2021. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1286721

ABSTRACT

Resumen Objetivo: Comparar la expresión de mRNA y proteínas de SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en pacientes con lesión intra-epitelial cervical de bajo y alto grado, con posterior progresión o regresión. Material y Método: Se realizó análisis de expresión de genes mediante RT-PCR y análisis de expresión de proteínas por inmunohistoquímica. El análisis estadís tico fue realizado con las pruebas: Wilcoxon, coeficiente de correlación de Spearman e índice de concordancia. Las muestras fueron pareadas en momento 1 y momento 2. Resultados: SFRP1 mostró tendencia de mayor expresión de mRNA en lesión intra-epitelial de bajo grado (momento 2) Vs. alto grado (momento 1). La expresión de proteínas por inmunohistoquímica de SFRP1 en casos de progresión (83,3 %) mostró disminución en su graduación (p = 0,0313*); los demás genes en estudio no tuvieron cambios estadísticamente significativos. Discusión: SFRP1 mostró comportamiento ajustado a resultados de estudios previos donde se encontró hipermetilado en lesiones intra-epiteliales de alto grado; su subexpresión por hipermetilación se reportó en otros canceres, proceso que colabora con su silenciamiento y transición epitelial-mesenquimatosa del cáncer de cuello uterino. Conclusiones. SFRP1 es potencial biomarcador en lesiones preneoplásicas del cuello uterino asociadas al virus de papiloma humano.


Abstract Objective. The aim of this work was to compare the expression of mRNA and proteins of SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 in patients with low and high grade cervical intraepithelial lesion, with subsequent progression or regression. Material and Methods: Gene expression analysis was conducted through RT-PCR and protein expression analysis was performed by immunohistochemistry. The statistics analysis were Wilcoxon test, Spearman's correlation coefficient and concordance index. The samples were paired during moment 1 (initial patient diag nosis) and moment 2 (follow-up histological diagnosis). Results: SFRP1 showed a trend of higher mRNA expression in low-grade intra-epithelial lesions (moment 2) Vs. high-grade (moment 1). The expression of proteins by immunohistochemistry of SFRP1 in progression cases (83.3%) showed a decrease in its graduation (p = 0.0313*); the other genes under study had no statistically significant. Discussion: SFRP1 showed a biological behavior adjusted to the results of previous studies where hypermethylation was found in high-grade intra-epithelial lesions; its subexpression by hypermethylation has been reported in other cancers, a process that collaborates with its silencing and epithelial-mesenchymal tran sition of cervical. Conclusions. SFRP1 is a potential biomarker in preneoplastic lesions of the cervix associated with human papillomavirus.


Subject(s)
Humans , Female , Adult , Papilloma , DNA Probes, HPV , Viruses , Proteins , Uterine Cervical Neoplasms , Disease Progression , Alphapapillomavirus
4.
Infectio ; 25(4): 241-249, oct.-dic. 2021. tab, graf
Article in English | LILACS, COLNAL | ID: biblio-1286717

ABSTRACT

Abstract Infection through the Hepatitis C virus does not have a vaccine and treatment with pegylated interferon and ribavirin can fail; which is why it may cause chronic infection and, consequently, could develop liver failure or hepatocellular carcinoma. It has been described that virus-cell recognition occurs between the E2 viral envelope protein and diverse cell receptors, with this interaction being critical in viral infection. which is why the study sought to identify inhibitory peptides of the interaction between viral E2 protein and the CD81 and CD209 receptors. Methodology: Through the RCSB protein database, crystals from the CD81 and CD209 receptors were selected, CD81/E2-HCV, CD209/E2-HCV complexes were carried out by SWISS-MODEL to generate inhibitory peptides of protein interaction through the Rosetta web server, this interaction was validated through ClusPro and finally, determined the theoretical physicochemical and cytotoxic properties of these peptides. Results: two peptides were obtained, without predicted toxicity, with a theoretical capacity of blocking the protein interaction between the E2 protein of the virus and CD81 and CD209.


Resumen La infección por el virus de la hepatitis C, no cuenta con vacuna y el tratamiento con interferón pegilado y ribavirina puede fallar; por lo que puede causar infec ción crónica y como consecuencia podría desarrollarse falla hepática o carcinoma hepatocelular. Se ha descrito que el reconocimiento virus-célula, se da entre la proteína de envoltura viral E2 y diversos receptores celulares, siendo esta interacción crítica en la infección viral. Razón por la cual este estudio buscó identificar péptidos inhibidores de la interacción entre la proteína E2 viral y los receptores CD81 y CD209. Metodología: A través de la base de datos de proteínas RCSB, se seleccionaron cristales de los receptores CD81 y CD209, se realizaron complejos CD81/E2-HCV, CD209/E2-HCV para generar péptidos inhibidores de interacción proteica a través del servidor web Rosetta, esta interacción fue validada a través de ClusPro y finalmente se evaluaron las propiedades fisicoquímicas y citotóxicas teóricas para estos péptidos. Resultados: se obtuvo dos péptidos, sin toxicidad predicha, con capacidad teórica de bloquear la interacción proteica entre la proteína E2 del virus y CD81 y CD209.


Subject(s)
Humans , Hepatitis Viruses , Peptides , Vaccines , Proteins , Hepatitis C , Liver Failure , Hepacivirus , Infections
5.
Rev. Investig. Salud. Univ. Boyacá ; 8(1): 75-90, 20210000. fig
Article in Spanish | LILACS, COLNAL | ID: biblio-1358961

ABSTRACT

Introducción: La babesiosis bovina es causada por parásitos Apicomplexa del género Babesia, siendo la Babesia bovis la especie asociada con cuadros clínicos más graves de la enfermedad. La invasión de B. bovis a los eritro-citos bovinos implica la interacción entre moléculas de los merozoítos del parásito con receptores de las células huésped. Por ende, conocer las proteínas involucradas en este proceso supone un importante paso para entender la biología del parásito. Objetivo: Describir las principales moléculas implicadas en el proceso de invasión de B. bovis a eritrocitos bovinos. Metodología: Se realizó una búsqueda en NCBI, Medline, LILACS y SciELO usando los términos: "Babesia bovis AND invasion process", "MSA-1", "RON2", "AMA-1", "moving junction", "B. bovis AND Vaccine candidates". Con corte en mayo de 2020, había 61 publicaciones disponibles en inglés que describen el estudio de las anteriores proteínas y su participación en la invasión.Resultados: Por ser clave el proceso de invasión a eritrocitos bovinos para la patogénesis de la babesiosis bovina, la revisión encontró 3 proteínas de B. bovis que participan en el reconocimiento e invasión a las células diana: MSA-1, AMA-1 y RON2. Sin embargo, los detalles a nivel molecular para las interacciones inter e intramoleculares aún no se han dilucidado por completo. Conclusiones: Conocer las moléculas involucradas en las interacciones parásito-hospedero permitirá entender cómo ocurre el proceso de invasión de B. bovis a los eritrocitos y, así, evaluar su futura utilidad como componente de una estrategia de control efectiva contra esta parasitosis


Introduction: Bovine babesiosis is caused by Apicomplexas parasites of the genus Babesia, Babesia bovis being the species associated with the most serious clinical conditions of the disease. B. bovisinvasion into the bovine erythrocytes involves the interaction between the parasites merozoites mo-lecules with host cell receptors. Therefore, knowing the proteins involved in the invasion process will enable understanding the parasite biology. Objective: To describe the important molecules involved in the B. bovis invasion process to bovine erythrocytes.Methodology: A search was made on NCBI, Medline, LILACS and SciELO databases using keywords as "Babesia bovis AND invasion process", "MSA-1", "RON2", "AMA-1", "moving junction", "B. bovis AND Vaccine candidates". 61 studies written in English describing the study for proteins that take place during invasion process which have been published until mayo were completely revised. Results: Given that the bovine erythrocyte invasion process is key for the pathogenesis of bovine babesiosis, a review was made where 3 proteins were found to be associated to the recognition and invasion processes of target cells: MSA-1, AMA-1 and RON2. However, the details at molecular level for the inter an intramolecular interaction have not yet been fully elucidated. Conclusions: Study the molecules involved in host-parasite interactions will allow understanding how the B. bovis invasion process to erythrocytes occurs and evaluating their future utility as a component of an effective control strategy for this parasitosis


Introdução: A babesiose bovina é causada por parasitas Apicomplexa do gênero Babesia, sendo a Babesia bovis a espécie associada com os sinais clínicos mais graves da doença. A invasão de B. bovis em eritrócitos bovinos envolve a interação entre moléculas dos merozoítos parasitas com receptores nas células hospedeiras. Por conseguinte, o conhecimento das proteínas envolvidas neste processo é um passo importante para a compreensão da biologia do parasita. Objetivo: Descrever as principais moléculas envolvidas no processo de invasão de B. bovis em eritró-citos bovinos. Metodologia: Foi realizada uma pesquisa no NCBI, Medline, LILACS e SciELO utilizando os termos: "Babesia bovis AND invasion process", "MSA-1", "RON2", "AMA-1", "moving junction", "B. bovis AND Vaccine candidates". Até maio de 2020 estavam disponíveis 61 publicações em inglês, que descreviam o estudo das proteínas acima referidas e o seu envolvimento na invasão. Resultados: Como o processo de invasão de eritrócitos bovinos é fundamental para á patogênese da babesiose bovina, a revisão encontrou 3 proteínas de B. bovis envolvidas no reconhecimento e invasão de células alvo: MSA-1, AMA-1 e RON2. No entanto, os detalhes a nível molecular para as interações Inter e intramoleculares ainda não foram completamente elucidados. Conclusões: A compreensão das moléculas envolvidas nas interações parasita-hospedeiro permitirá entender como ocorre o processo da invasão de B. bovis em eritrócitos e, assim, avaliar sua utilidade futura como componente de uma estratégia efetiva de controle contra esta parasitose


Subject(s)
Babesia bovis , Babesiosis , Proteins , Infection Control , Host-Parasite Interactions
6.
Braz. J. Pharm. Sci. (Online) ; 57: e19061, 2021. tab, graf
Article in English | LILACS | ID: biblio-1350245

ABSTRACT

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.


Subject(s)
Peptides/pharmacology , Proteins/pharmacology , Soybean Proteins/pharmacology , Dietary Supplements , Protein Hydrolysates/pharmacology , Mass Spectrometry , Chromatography, Liquid/methods , Process Optimization/classification , Hydrogen-Ion Concentration , Hypertension/therapy , Antihypertensive Agents/analysis
8.
Article in English | WPRIM | ID: wpr-888783

ABSTRACT

A large number of protease inhibitors have been found from leeches, which are essential in various physiological and biological processes. In the curret study, a novel elastase inhibitor was purified and characterized from the leech of Hirudinaria manillensis, which was named HMEI-A. Primary structure analysis showed that HMEI-A belonged to a new family of proteins. HMEI-A exerted inhibitory effects on elastase and showed potent abilities to inhibit elastase with an inhibition constant (K


Subject(s)
Amino Acid Sequence , Animals , Leeches/chemistry , Pancreatic Elastase/antagonists & inhibitors , Protease Inhibitors/pharmacology , Proteins
9.
Frontiers of Medicine ; (4): 608-620, 2021.
Article in English | WPRIM | ID: wpr-888743

ABSTRACT

t(8;21)(q22;q22) acute myeloid leukemia (AML) is a highly heterogeneous hematological malignancy with a high relapse rate in China. Two leukemic myeloblast populations (CD34


Subject(s)
Gene Expression , Granulocyte Precursor Cells , Humans , Immunophenotyping , Leukemia, Myeloid, Acute/genetics , Membrane Glycoproteins , Prognosis , Proteins , Proto-Oncogene Proteins c-kit/genetics
10.
Frontiers of Medicine ; (4): 541-550, 2021.
Article in English | WPRIM | ID: wpr-888740

ABSTRACT

Synthetic lethal screening, which exploits the combination of mutations that result in cell death, is a promising method for identifying novel drug targets. This method provides a new avenue for targeting "undruggable" proteins, such as c-Myc. Here, we revisit current methods used to target c-Myc and discuss the important functional nodes related to c-Myc in non-oncogene addicted network, whose inhibition may cause a catastrophe for tumor cell destiny but not for normal cells. We further discuss strategies to identify these functional nodes in the context of synthetic lethality. We review the progress and shortcomings of this research field and look forward to opportunities offered by synthetic lethal screening to treat tumors potently.


Subject(s)
Humans , Mutation , Neoplasms/genetics , Proteins , Proto-Oncogene Proteins c-myc/genetics , Synthetic Lethal Mutations
11.
Article in Chinese | WPRIM | ID: wpr-888362

ABSTRACT

OBJECTIVE@#To screen proteins interacting with ring finger protein 216(RNF216) through yeast two hybrid experiment, and further clarify the role of RNF216 in the pathogenesis of gonadotropin-releasing hormone deficiency.@*METHODS@#A recombinant expression vector pGBKT7-RNF216 was constructed and transformed into yeast Y2HGold, which was hybridized with a human cDNA library in order to screen proteins interacting with RNF216. The interaction was verified in yeast Y2HGold.@*RESULTS@#A recombinant expression vector pGBKT7-RNF216 was successfully constructed and expressed in yeast Y2HGold. Filamin B (FLNB) was identified by yeast two hybrid experiment, and their interaction was verified in yeast Y2HGold.@*CONCLUSION@#An interaction between FLNB and RNF216 was identified through yeast two hybrid experiment. RNF216 may affect the proliferation and migration of GnRH neurons by regulating FLNB or FLNB/FLNA heterodimers.


Subject(s)
Gene Library , Gonadotropin-Releasing Hormone/genetics , Humans , Proteins , Two-Hybrid System Techniques , Ubiquitin-Protein Ligases/genetics
12.
Braz. J. Vet. Res. Anim. Sci. (Online) ; 58(n.esp): e174301, 2021. tab
Article in English | LILACS, VETINDEX | ID: biblio-1348265

ABSTRACT

Developing effective cooled semen protocols is essential to increase pregnancy rates and reproductive efficiency in donkeys. This study aimed to evaluate the effect on sperm kinetic parameters and membrane integrity in cooled donkey semen diluted with defined milk proteins extender with 1% or 2% of egg yolk and the removal of seminal plasma. Twenty-four ejaculates from six jackasses were collected. Each ejaculate was divided into four aliquots that were diluted in extender with 1% (EY1) or 2% (EY2) egg yolk. One sample from each group was centrifuged, seminal plasma was removed (CEY1, CEY2 groups, respectively), and the samples were then refrigerated at 5 °C for 24 h. Fresh and cooled semen samples were assessed for sperm motility, morphology, and plasma membrane integrity. Total motility, progressive motility, sperm kinetic parameters, or live sperm cells were not statistically different when semen was cooled with an extender supplemented with 1% or 2% of egg yolk. Seminal plasma removal does not affect total motility or sperm kinetic parameters. However, progressive motility decreased (P<0.05) when semen was extended with 2% of egg yolk and seminal plasma was removed. Membrane integrity was affected (P<0.05) in centrifuged samples. In conclusion, the obtained results suggest that there is no difference in sperm kinetics and membrane integrity when 1% or 2% of egg yolk was added to the Equiplus extender. Also, the removal of seminal plasma by centrifugation did not have any beneficial effect on cooled donkey semen. Further studies are needed to relate these results with in vivo fertility tests with cooled donkey semen.(AU)


O desenvolvimento de protocolos de sêmen resfriado eficazes é essencial para aumentar as taxas de prenhez e eficiência reprodutiva em jumentos. O objetivo desse estudo foi avaliar o efeito do diluente à base de proteínas do leite com 1 ou 2% de gema de ovo sobre os parâmetros cinéticos do sêmen e integridade da membrana em sêmen resfriado de jumento, com ou sem a remoção do plasma seminal. Vinte e quatro ejaculados de seis jumentos foram coletados. Cada ejaculado foi dividido em quatro alíquotas e diluído em diluente com 1% (EY1) ou 2% (EY2) de gema de ovo. Uma amostra por grupo foi centrifugada e o plasma seminal removido (grupos CEY1 e CEY2, respectivamente). Os pellets foram novamente ressuspendidos nas mesmas concentrações e diluentes. Em seguida, as quatro alíquotas foram refrigeradas a 5°C por 24 horas. Amostras de sêmen fresco e refrigerado foram avaliadas quanto à motilidade espermática e integridade da membrana plasmática. Motilidade total, motilidade progressiva, parâmetros de cinética espermática ou células espermáticas vivas não apresentaram diferença significativa quando o sêmen foi resfriado com diluente suplementado com 1% ou 2% de gema de ovo. A remoção do plasma seminal não afetou a motilidade total ou os parâmetros de cinética espermática; entretanto, a motilidade progressiva diminuiu (P<0,05) quando o sêmen foi diluído com 2% de gema de ovo e o plasma seminal removido. Nas amostras centrifugadas, a integridade da membrana foi afetada (P<0,05). Em conclusão, os resultados sugerem que não há diferença na cinética espermática e na integridade da membrana quando 1% ou 2% de gema de ovo são adicionados ao diluente Equiplus e a remoção do plasma seminal por centrifugação não teve nenhum efeito benéfico no resfriamento de sêmen de jumento. Mais estudos são necessários para relacionar esses resultados com testes de fertilidade in vivo com sêmen resfriado em jumentos.(AU)


Subject(s)
Animals , Plasma , Semen Preservation/veterinary , Sperm Motility , Cryopreservation , Equidae , Egg Yolk , Semen , Proteins
13.
Chinese Journal of Biotechnology ; (12): 1131-1138, 2021.
Article in Chinese | WPRIM | ID: wpr-878619

ABSTRACT

Identification of the target proteins of small molecule drugs is crucial for understanding the mechanisms of drug actions and its side effects. Conventional methods require chemical modification, which might alter the activities of the drugs. Various label-free techniques have been developed to identify drug target proteins without chemical modifications. This includes drug affinity responsive target stability (DARTS), stability of proteins from rates of oxidation (SPROX), cellular thermal shift assay (CETSA), thermal proteome profiling (TPP) and many others. Here we review the principles and applications of these label-free techniques, their advantages and limitations, as well as the most recent advances.


Subject(s)
Drug Delivery Systems , Pharmaceutical Preparations , Proteins
14.
Chinese Journal of Biotechnology ; (12): 561-579, 2021.
Article in Chinese | WPRIM | ID: wpr-878582

ABSTRACT

Proteases are widely found in organisms participating in the decomposition of proteins to maintain the organisms' normal life activities. Protease inhibitors regulate the activities of target proteases by binding to their active sites, thereby affecting protein metabolism. The key amino acid mutations in proteases and protease inhibitors can affect their physiological functions, stability, catalytic activity, and inhibition specificity. More active, stable, specific, environmentally friendly and cheap proteases and protease inhibitors might be obtained by excavating various natural mutants of proteases and protease inhibitors, analyzing their key active sites by using protein engineering methods. Here, we review the studies on proteases' key active sites and protease inhibitors to deepen the understanding of the active mechanism of proteases and their inhibitors.


Subject(s)
Binding Sites , Catalytic Domain , Endopeptidases , Peptide Hydrolases/genetics , Protease Inhibitors , Proteins
15.
Chinese Journal of Biotechnology ; (12): 486-499, 2021.
Article in Chinese | WPRIM | ID: wpr-878577

ABSTRACT

Laboratory evolution is an important approach to improve the performance of microorganisms. In the past decades, the methods for laboratory evolution have developed rapidly and applied widely. However, the commonly used evolution strategies for strains or specific proteins cannot achieve continuous mutation, and require multiple rounds of operation, therefore they are considered as a labor intensive process. The development of mutation and screening technologies have facilitated the development of continuous evolution in vivo and greatly improved the efficiency of laboratory evolution. The continuous in vivo evolution achieves in vivo mutation, perfectly combining mutation with screening to evolve a specific phenotype with minimal human intervention. This review summarizes the recent advances of in vivo continuous evolution technologies for either genome-scale mutation or evolution of specific proteins. The principles of these technologies and their applications are introduced. On this basis, the advantages and limitations of these technologies are discussed. We also give a perspective of future development of continuous in vivo evolution.


Subject(s)
Directed Molecular Evolution , Humans , Mutation , Phenotype , Proteins
16.
Chinese Journal of Biotechnology ; (12): 163-177, 2021.
Article in Chinese | WPRIM | ID: wpr-878551

ABSTRACT

Directed evolution is a cyclic process that alternates between constructing different genes and screening functional gene variants. It has been widely used in optimization and analysis of DNA sequence, gene function and protein structure. It includes random gene libraries construction, gene expression in suitable hosts and mutant libraries screening. The key to construct gene library is the storage capacity and mutation diversity, to screen is high sensitivity and high throughput. This review discusses the latest advances in directed evolution. These new technologies greatly accelerate and simplify the traditional directional evolution process and promote the development of directed evolution.


Subject(s)
Base Sequence , Directed Molecular Evolution , Gene Library , Mutation , Proteins/genetics
17.
Chinese Journal of Biotechnology ; (12): 4201-4214, 2021.
Article in Chinese | WPRIM | ID: wpr-921499

ABSTRACT

Protein kinase CK2 is a common, evolutionarily conserved and ubiquitous protein kinase. In recent years, increasing evidences have shown that CK2 has a variety of phosphorylated protein substrates, which play important roles in growth, development and various diseases. Therefore, CK2 may participate in these physiological processes by regulating the phosphorylation of these substrates. This article briefly reviewed the structural characteristics of protein kinase CK2 and its physiological functions in growth, development, immunity, formation of tumor and other diseases, in order to provide knowledge basis for further research on the regulatory mechanism of CK2 and potential applications of its inhibitors.


Subject(s)
Casein Kinase II/metabolism , Phosphorylation , Proteins
18.
Chinese Journal of Biotechnology ; (12): 4187-4200, 2021.
Article in Chinese | WPRIM | ID: wpr-921498

ABSTRACT

The amino acid sequence of ancestral enzymes from extinct organisms can be deduced through in silico approach termed ancestral sequence reconstruction (ASR). ASR usually has six steps, which are the collection of nucleic acid/amino acid sequences of modern enzymes, multiple sequence alignment, phylogenetic tree construction, computational deduction of ancestral enzyme sequence, gene cloning, and characterization of enzyme properties. This method is widely used to study the adaptation and evolution mechanism of molecules to the changing environmental conditions on planetary time scale. As enzymes play key roles in biocatalysis, this method has become a powerful method for studying the relationship among the sequence, structure, and function of enzymes. Notably, most of the ancestral enzymes show better temperature stability and mutation stability, making them ideal protein scaffolds for further directed evolution. This article summarizes the computer algorithms, applications, and commonly used computer software of ASR, and discusses the potential application in directed evolution of enzymes.


Subject(s)
Amino Acid Sequence , Evolution, Molecular , Phylogeny , Proteins/genetics , Sequence Alignment
19.
Chinese Journal of Biotechnology ; (12): 3915-3932, 2021.
Article in Chinese | WPRIM | ID: wpr-921476

ABSTRACT

Targeted protein degradation (TPD) technology facilitates specific and efficient degradation of disease-related proteins through hijacking the two major protein degradation systems in mammalian cells: ubiquitin-proteasome system and lysosome pathway. Compared with traditional small molecule-inhibitors, TPD-based drugs exhibit the characteristics of a broader target spectrum. Compared with techniques interfere with protein expression on the gene and mRNA level, TPD-based drugs are target-specific, efficaciously rapid, and not constrained by post-translational modification of proteins. In the past 20 years, various TPD-based technologies have been developed. Most excitingly, two TPD-based therapeutic drugs have been approved by FDA for phase Ⅰ clinical trials in 2019. Despite of the early stage characteristics and various obstructions of the TPD technology, it could serve as a powerful tool for the development of novel drugs. This review summarizes the advances of different degradation systems based on TPD technologies and their applications in disease therapy. Moreover, the advantages and challenges of various technologies were discussed systematically, with the aim to provide theoretical guidance for further application of TPD technologies in scientific research and drug development.


Subject(s)
Animals , Proteasome Endopeptidase Complex/metabolism , Protein Processing, Post-Translational , Proteins/metabolism , Proteolysis , Technology
20.
Chinese Journal of Biotechnology ; (12): 3863-3879, 2021.
Article in Chinese | WPRIM | ID: wpr-921472

ABSTRACT

The accumulation of protein sequence and structure data allows researchers to obtain large amount of descriptive information, simultaneously it poses an urgent need for researchers to extract information from existing data efficiently and apply it to downstream tasks. Protein design enables the development of novel proteins that are no longer restricted by experimental conditions, which is of great significance for drug target prediction, drug discovery, and material design. As an efficient method for data feature extraction, deep learning can be used to model protein data, and further add a priori information to design novel proteins. Therefore, protein design based on deep learning has become a promising approach despite of many challenges. This review summarizes the deep learning-based modeling and design methods of protein sequence and structure data, highlighting the strategies, principle, scope of application and case studies, with the aim to provide a valuable reference for relevant researchers.


Subject(s)
Amino Acid Sequence , Deep Learning , Drug Development , Proteins
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