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1.
Int J Biol Macromol ; 194: 800-810, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34848239

RESUMO

The hydrophobic nature of wool induced by its surface lipid barrier hinders its wettability during processing. Scouring of wool is conducted to remove this lipid barrier and facilitate any wet processes. Scouring of wool is conducted using soda ash followed by rinsing with huge amount of water to ensure complete removal of alkali. This work aimed at utilization of thermophilic lipase enzyme for removal of wool surface lipid barrier without deterioration on the fibre interior. A thermally stable lipase enzyme was produced from thermophilic microorganism; namely Bacillus aryabhattai B8W22, and was utilized in bio-scouring of wool. The produced enzyme was immobilized on sericin-based discs to enhance its stability and to make it reusable. The activity of both free and immobilized lipase enzymes at different conditions was assessed. The effects of bio-scouring of wool on its dyeability with acid, basic, and reactive dyes, as well as on some of its inherent properties, were monitored. Results showed that the bio-scoured wool exhibits enhanced dyeability with the said classes of dyes more than that of conventionally scoured samples. One-bath scouring and dyeing of wool fibres in two successive steps was conducted to reduce consumption of water and energy during wet processing of wool.


Assuntos
Enzimas Imobilizadas , Lipase/química , Fibra de Lã/análise , Lã/química , Animais , Bacillus/classificação , Bacillus/enzimologia , Bacillus/genética , Proteínas de Bactérias/química , Corantes/química , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Lipólise , Estrutura Molecular , Temperatura
2.
Molecules ; 26(15)2021 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-34361733

RESUMO

Biocomposites comprising a combination of natural fibres and bio-based polymers are good alternatives to those produced from synthetic components in terms of sustainability and environmental issues. However, it is well known that water or aqueous chemical solutions affect natural polymers/fibres more than the respective synthetic components. In this study the effects of water, salt water, acidic and alkali solutions ageing on water uptake, mechanical properties and flammability of natural fibre-reinforced polypropylene (PP) and poly(lactic acid) (PLA) composites were compared. Jute, sisal and wool fibre- reinforced PP and PLA composites were prepared using a novel, patented nonwoven technology followed by the hot press method. The prepared composites were aged in water and chemical solutions for up to 3 week periods. Water absorption, flexural properties and the thermal and flammability performances of the composites were investigated before and after ageing each process. The effect of post-ageing drying on the retention of mechanical and flammability properties has also been studied. A linear relationship between irreversible flexural modulus reduction and water adsorption/desorption was observed. The aqueous chemical solutions caused further but minor effects in terms of moisture sorption and flexural modulus changes. PLA composites were affected more than the respective PP composites, because of their hydrolytic sensitivity. From thermal analytical results, these changes in PP composites could be attributed to ageing effects on fibres, whereas in PLA composite changes related to both those of fibres present and of the polymer. Ageing however, had no adverse effect on the flammability of the composites.


Assuntos
Materiais Biocompatíveis/química , Retardadores de Chama/análise , Fibras Minerais/análise , Poliésteres/química , Polipropilenos/química , Água/química , Materiais Biocompatíveis/análise , Fibra de Algodão/análise , Temperatura Alta , Humanos , Teste de Materiais , Poliésteres/análise , Polipropilenos/análise , Soluções , Fibra de Lã/análise
3.
Int J Biol Macromol ; 181: 291-300, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-33775760

RESUMO

The purpose of this work was to evaluate the effect of itaconation on sizing properties (such as viscosity stability, adhesion and film properties) of biological macromolecule (corn starch) for developing a new bio-based sizing agent [itaconylated starch (IS)]. Granular IS samples were characterized by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) techniques. The adhesion of IS to wool fibers was investigated by a standard method (FZ/T 15001-2008). And film properties of IS samples were also studied in terms of tensile strength, breaking elongation, bending endurance and degree of crystallinity, etc. Compared with control acid-converted starch (ACS), stronger bonding forces to wool fibers for IS as well as higher breaking elongation and lower tensile strength for IS film were displayed. Increasing the degrees of substitution (DS) of IS samples from 0 to 0.052 was able to achieve gradually enhanced bonding forces, breaking elongation and bending endurance, which implied that increasing the number of itaconate substituents could play a significantly positive role in overcoming the shortcomings (insufficient adhesion and film brittleness) of starch. These experimental results denoted that the granular IS exhibited potential for the use as a new starch-based size in the sizing of wool warp yarns.


Assuntos
Amido/química , Succinatos/química , Fibra de Lã/análise , Ácidos/química , Adesividade , Espectroscopia de Infravermelho com Transformada de Fourier , Amido/ultraestrutura , Tensão Superficial , Resistência à Tração , Viscosidade , Molhabilidade , Difração de Raios X
4.
Proteins ; 89(6): 708-720, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33550642

RESUMO

Trichocyte keratin intermediate filament proteins (keratins) and keratin associated proteins (KAPs) differ from their epithelial equivalents by having significantly more cysteine residues. Interactions between these cysteine residues within a mammalian fiber, and the putative regular organization of interactions are likely important for defining fiber mechanical properties, and thus biological functionality of hairs. Here we extend a previous study of cysteine accessibility under different levels of exposure to reducing compounds to detect a greater resolution of statistically non-random interactions between individual residues from keratins and KAPs. We found that most of the cysteines with this non-random accessibility in the KAPs were close to either the N- or C- terminal domains of these proteins. The most accessible non-random cysteines in keratins were present in the head or tail domains, indicating the likely function of cysteine residues in these regions is in readily forming intermolecular bonds with KAPs. Some of the less accessible non-random cysteines in keratins were discovered either close to or within the rod region in positions previously identified in human epithelial keratins as involved in crosslinking between the heterodimers of the tetramer. Our present study therefore provides a deeper understanding of the accessibility of disulfides in both keratins and KAPs and thus proves that there is some specificity to the disulfide bond interactions leading to these inter- and intra-molecular bonds stabilizing the fiber structure. Furthermore, these suggest potential sites of interaction between keratins and KAPs as well as keratin-keratin interactions in the trichocyte intermediate filament.


Assuntos
Cisteína/química , Dissulfetos/química , Queratinas Específicas do Cabelo/química , Mapeamento de Peptídeos/métodos , Fibra de Lã/análise , Acrilamida/química , Alquilação , Sequência de Aminoácidos , Animais , Cromatografia Líquida , Humanos , Iodoacetamida/química , Ácido Iodoacético/química , Queratinas Específicas do Cabelo/classificação , Isoformas de Proteínas/química , Isoformas de Proteínas/classificação , Multimerização Proteica , Carneiro Doméstico , Espectrometria de Massas em Tandem , Lã/química
5.
Int J Legal Med ; 135(5): 1709-1715, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33587179

RESUMO

When dealing with complex crimes such as rape and assault, every trace takes on an essential role. The hands are often the only means of defence and offence for the victim as well as a frequent area of contact with the environment; fingernails of a victim are a well-known possible source of DNA of the aggressor; nevertheless, they are more rarely treated as an area of interest for non-genetic material, particularly on living victims. The hyponychium, because of its physiological protective function, lends itself ideally to retaining different kinds of traces representative of an environment or various products and substrates that could shed light on the environment and objects involved in the event. We therefore tested how far this capability of the hyponychium could go by simulating the dynamics of contamination of the nail through scratching on different substrates (brick and mortar, painted wood, ivy leaves, cotton and woollen fabric, soil) and persistence of any contaminant at different time intervals. We have thus shown how these traces may remain in the living for up to 24 h after the event using inexpensive and non-destructive techniques such as the episcopic and optical microscope.


Assuntos
Vítimas de Crime , Ciências Forenses , Microscopia , Unhas/química , Manejo de Espécimes/métodos , Fibra de Algodão/análise , Hedera , Humanos , Pintura/análise , Projetos Piloto , Solo , Madeira/análise , Fibra de Lã/análise
6.
Genes (Basel) ; 11(8)2020 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-32823629

RESUMO

Variation in some caprine keratin-associated protein (KAP) genes has been associated with cashmere fiber traits, but many KAP genes remain unidentified in goats. In this study, we confirm the identification of a KAP27-1 gene (KRTAP27-1) and describe its effect on cashmere traits in 248 Longdong cashmere goats. A polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis was used to screen for sequence variation in this gene, and three sequence variants (named A to C) were found. These sequences have the highest similarity (77% identity) to a human KRTAP27-1 sequence, while sharing some homology with a predicted caprine KRTAP27-1 sequence ENSCHIG00000023347 in the goat genome construct (ARS1:CM004562.1) at chromosome 1 position 3,966,193-3,973,677 in the forward strand. There were two single nucleotide polymorphisms (SNPs) detected in the coding sequence, including one nonsynonymous SNP (c.413C/T; p.Ala138Val) and one synonymous SNP (c.495C/T). The C variant differed from A and B at c.413C/T, having cytosine in its nucleotide sequence, while the B variant differed from A and C at c.495C/T, having thymine in its nucleotide sequence. Goats of the genotypes AB and BB produced cashmere fibers of higher mean fiber diameter (MFD) than goats of genotype AA, but no difference in MFD was detected between the AB and BB goats. These results suggest that B is associated with increased MFD. Expression of the caprine KRTAP27-1 sequence was predominantly detected in the skin tissue of goats but not or only weakly detected in other tissues, including longissimus dorsi muscle, heart, kidney, liver, lung and spleen.


Assuntos
Estudos de Associação Genética , Cabras/genética , Queratinas/genética , Característica Quantitativa Herdável , Fibra de Lã/análise , Animais , Sequência de Bases , Expressão Gênica , Queratinas/metabolismo , Fenótipo , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas
7.
Arq. bras. med. vet. zootec. (Online) ; 72(3): 977-984, May-June, 2020. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-1129703

RESUMO

O objetivo deste estudo foi avaliar o efeito da suplementação durante diferentes períodos gestacionais sobre a produção de lã de ovelhas e cordeiros da raça Ideal. Foram utilizadas 53 ovelhas da raça Ideal, com escore corporal médio (3), inseminadas pela técnica de laparoscopia, com sêmen fresco de um único reprodutor; e seus cordeiros. Os tratamentos experimentais foram: sem suplementação (n=9); suplementação do início da gestação até 50 dias (n=11); suplementação dos 51 aos 100 dias de gestação (n=11); suplementação dos 101 aos 150 dias - final da gestação (n=11); e suplementação durante toda a gestação (n=11). A suplementação foi a 1,5% do peso corporal. Amostras de lã das ovelhas e dos cordeiros foram tomadas na região do costilhar esquerdo e enviadas ao laboratório para análises objetivas de finura de lã. As suplementações no terço final e durante toda a gestação proporcionavam os melhores resultados, com aumento de produção de lã e de peso das ovelhas. Ovelhas que receberam suplementação durante toda a gestação apresentaram maior diâmetro de fibra e peso corporal. O desempenho de lã dos cordeiros não foi influenciado pela alimentação de ovelhas durante a gestação.(AU)


The objective of this study was to evaluate the effect of supplementation during different gestational periods on the wool production of sheep and lambs of the Ideal breed. Fifty-three adult Ideal sheep were used, with a mean body score inseminated by the laparoscopy technique using fresh sperm from a single breeder and his lambs. Treatment groups: No supplementation (n=9); Supplementation in the beginning of gestation up to 50 days (n=11); Supplementation from 51 to 100 days of gestation (n=11); Supplementation from 101 to 150 days - end of gestation (n=11) ;and Supplementation throughout the gestation (n=11). The supplementation was at 1.5% of body weight. After birth, lambs were kept with their mothers in cultivated pasture of black oats and ryegrass. Wool samples from sheep and lambs were taken in the left-hand region and sent to the laboratory for objective analyzes of wool fineness. The supplementation performed in the final third and throughout the gestation provides the best results, with increased sheep weight. Ewes that received supplementation throughout pregnancy had greater fiber diameter and body weight. The wool performance of lambs was not influenced by feeding sheep during gestation.(AU)


Assuntos
Animais , , Peso Corporal , Ovinos , Suplementos Nutricionais , Fibra de Lã/análise
8.
PLoS One ; 15(3): e0228332, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32160199

RESUMO

Management of camelids in the coastal valleys of the Andes has generated much debate in recent years. Zooarchaeological and isotopic studies have demonstrated that in the coastal valleys of northern and southern Peru there were locally maintained camelid herds. Because of the hyperarid conditions of the northern coast of Chile, this region has been assumed to be unsuitable for the raising of camelids. In this study we report stable carbon and nitrogen isotopic compositions of camelid bone collagen and textiles made from camelid fiber from Late Intermediate Period (LIP) and Late Horizon (LH) occupations in northern Chilean river valleys. The camelid bone collagen isotopic compositions are consistent with these animals originating in the highlands, although there is a significant difference in the camelids dating to the LIP and LH, possibly because of changes made to distribution and exchange networks by the Inca in the LH. There were no differences between the isotopic compositions of the camelid fibers sampled from textiles in the LIP and LH, suggesting that either the production of camelid fiber was unchanged by the Inca or the changes that were made do not present visible isotopic evidence. Several camelid fiber samples from both the LIP and LH present very high δ13C and δ15N values, comparable to human hair samples from one site (Huancarane) in the Camarones Valley. These data suggest that people in the northern valleys of Chile may have kept small numbers of animals specifically for fiber production. Overall, however, the vast majority of the textile samples have isotopic compositions that are consistent with an origin in the highlands. These data suggest that the hyperarid coastal river valleys of northern Chile did not support substantial camelid herds as has been interpreted for northern Peru.


Assuntos
Criação de Animais Domésticos/história , Camelídeos Americanos , Fibra de Lã/história , Animais , Arqueologia , Osso e Ossos/química , Isótopos de Carbono/análise , Chile , Colágeno/química , Fósseis , História do Século XV , História do Século XVI , História Medieval , Isótopos de Nitrogênio/análise , Peru , Fibra de Lã/análise
9.
Genes (Basel) ; 11(2)2020 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-32019077

RESUMO

Wool and hair fibres consist of a variety of proteins, including the keratin-associated proteins (KAPs). In this study, a putative ovine homologue of the human KAP21-2 gene (KRTAP21-2) was identified. It was located on chromosome 1 as a 201-bp open reading frame (ORF) in the ovine genome assembly from a Texel sheep (v.4 NC_019458.2: nt122932727 to 122932927). A polymerase chain reaction- single strand conformation polymorphism (PCR-SSCP) analysis of this ORF, and subsequent DNA sequencing, identified five sequences (named A-E). The putative amino acid sequences that would be produced, shared some identity with each other and with other KAPs, but they were most similar to ovine KAP21-1, and phylogenetically related to human KAP21-2. The location of the ovine KRTAP21-2 sequence was consistent with the location of human KRTAP21-2, and this suggests they represent different variant forms of ovine KRTAP21-2. Variation in this gene was investigated in 389 Merino (sire) × Southdown-cross (ewe) lambs. These were derived from four independent sire-lines. The sequence variation was found to be associated with variation in five wool traits: including mean staple length (MSL), mean fibre diameter (MFD), fibre diameter standard deviation (FDSD), prickle factor (PF), and greasy fleece weight (GFW). The most persistent effect of KRTAP21-2 variation was with variation in MSL; with the MSL of sheep of genotype AC being 12.5% greater than those of genotype CE. A similar effect was observed from individual variant absence/presence models. This suggests that KRTAP21-2 should be further investigated as a possible gene-marker for improving MSL.


Assuntos
Queratinas/genética , Análise de Sequência de DNA/métodos , Ovinos/genética , Fibra de Lã/análise , Animais , Mapeamento Cromossômico , Cromossomos de Mamíferos/genética , Filogenia , Homologia de Sequência do Ácido Nucleico
10.
Biosens Bioelectron ; 141: 111480, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31272056

RESUMO

The technological quest for flexible devices to be interfaced with the biological world has driven the recent reinvention of bioderived polymers as multifunctional active and passive constituent elements for electronic and photonic devices to use in the biomedical field. Keratin is one of the most important structural proteins in nature to be used as biomaterial platform in view of the recently reported advances in the extraction and processing from hair and wool fibers. In this article we report for the first time the simultaneous use of naturally extracted keratin as both active ionic electrolyte for water ions sensing and as bendable and insoluble substrate into the same multielectrode array-based device. We implemented the multifunctional system exclusively made by keratin as a bendable sensor for monitoring the humidity flow. The enhancement of the functional and structural properties of keratin such as bendability and insolubility were obtained by unprecedented selective chemical doping. The mechanisms at the basis of the sensing of humidity in the device were investigated by cyclic voltammetry and rationalized by reversible binding and extraction of water ions from the volume of the keratin active layer, while the figures of merit of the biopolymer such as the ionic conductivity and relaxation time were determined by means of electrical impedance and dielectric relaxation spectroscopy. A reliable linear correlation between the controlled-humidity level and the amperometric output signal together with the assessment on measure variance are demonstrated. Collectively, the fine-tuned ionic-electrical characterization and the validation in controlled conditions of the free-standing insoluble all-keratin made microelectrode array ionic sensor pave the way for the effective use of keratin biopolymer in wearable or edible electronics where conformability, reliability and biocompatibility are key-enabling features.


Assuntos
Técnicas Biossensoriais/instrumentação , Umidade , Queratinas/química , Vapor/análise , Dispositivos Eletrônicos Vestíveis , Animais , Materiais Biocompatíveis/química , Eletricidade , Microeletrodos , Fibra de Lã/análise
11.
Molecules ; 24(12)2019 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-31234558

RESUMO

Natural tannins from various plants have been used throughout human history in textile dyeing, often as mordant dyes. The ageing behavior of these dyes is a challenge in conservation science, requiring a thorough knowledge of the textile-mordant-dye system. In this work, we analyzed reference wool yarns dyed with natural tannins from oak gallnuts, walnut (Juglans regia), and catechu (Acacia catechu), after artificial ageing. To gain insights on the composition of the dyestuffs and on how they aged, an analytical procedure based on extraction with Na2EDTA/DMF (ethylenediaminetetraacetic acid/dimethylformamide) and high-performance liquid chromatography (HPLC) analysis using high-resolution mass spectrometry detection was used. Since conventional reversed-phase (RP) columns usually show poor retention efficiency of highly polar compounds such as tannins, an RP-amide embedded polar group stationary phase was used to achieve optimal retention of the most polar compounds. Tannins from oak gallnuts showed little degradation after ageing, while a significant increase in the content of hydroxybenzoic acids was observed for tannins from walnut and catechu. Finally, the analytical procedure was applied to characterize the tannin dyes in historical tapestries from the 15th to 16th century, and the results were discussed in comparison with the reference yarns.


Assuntos
Espectrometria de Massas , Taninos/análise , Taninos/química , Fibra de Lã/análise , Animais , Juglans/química , Espectrometria de Massas/métodos , Estrutura Molecular , Peso Molecular , Nozes/química , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/química
12.
Molecules ; 24(3)2019 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-30754712

RESUMO

The aqueous extract of dry onion skin waste from the 'Dorata di Parma' cultivar was tested as a new source of biomolecules for the production of colored and biofunctional wool yarns, through environmentally friendly dyeing procedures. Specific attention was paid to the antioxidant and UV protection properties of the resulting textiles. On the basis of spectrophotometric and mass spectrometry analyses, the obtained deep red-brown color was assigned to quercetin and its glycoside derivatives. The Folin⁻Ciocalteu method revealed good phenol uptakes on the wool fiber (higher than 27% for the textile after the first dyeing cycle), with respect to the original total content estimated in the water extract (78.50 ± 2.49 mg equivalent gallic acid/g onion skin). The manufactured materials showed remarkable antioxidant activity and ability to protect human skin against lipid peroxidation following UV radiation: 7.65 ± 1.43 (FRAP assay) and 13.60 (ORAC assay) mg equivalent trolox/g textile; lipid peroxidation inhibition up to 89.37%. This photoprotective and antioxidant activity were therefore ascribed to the polyphenol pool contained in the outer dried gold skins of onion. It is worth noting that citofluorimetric analysis demonstrated that the aqueous extract does not have a significative influence on cell viability, neither is capable of inducing a proapoptotic effect.


Assuntos
Antioxidantes/farmacologia , Cebolas/química , Polifenóis/farmacologia , Protetores contra Radiação/farmacologia , Pele/efeitos dos fármacos , Fibra de Lã/análise , Animais , Antioxidantes/química , Sobrevivência Celular , Ácido Gálico , Glicosídeos/química , Glicosídeos/farmacologia , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos da radiação , Espectrometria de Massas , Camundongos , Extratos Vegetais/química , Polifenóis/química , Quercetina/análogos & derivados , Quercetina/química , Células RAW 264.7 , Protetores contra Radiação/química , Pele/efeitos da radiação , Espectrofotometria , Indústria Têxtil
13.
Int J Biol Macromol ; 125: 1008-1015, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30572050

RESUMO

Water soluble keratose proteins were obtained from an Ovis Aries wool using peracetic acid oxidation. The wool samples and the extracted keratose proteins were characterized by using FTIR, XRD, SEM and TGA techniques. Fractions of α-keratose (MW = 43-53 kDa) along with protein species with molecular weights between 23 kDa and 33 kDa were identified in the SDS-PAGE analysis result of the extracted protein mixture. DLS and AFM experiments indicated that self-assembled globular nanoparticles with diameters between 15 nm and 100 nm formed at 5 mg/ml keratose concentration. On the other hand, upon incubation of 10 w % keratose solutions at 37 °C and 50 °C, interconnected keratose hydrogels with respective storage modulus (G') values of 0.17 ±â€¯0.03 kPa and 3.7 ±â€¯0.5 kPa were obtained. It was shown that the keratose hydrogel prepared at 37 °C supported L929 mouse fibroblast cell proliferation which suggested that these keratose hydrogels could be promising candidates in soft tissue engineering applications.


Assuntos
Materiais Biocompatíveis/química , Hidrogéis/química , Queratinas/isolamento & purificação , Nanopartículas/química , Alicerces Teciduais , Fibra de Lã/análise , Animais , Materiais Biocompatíveis/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Hidrogéis/farmacologia , Queratinas/farmacologia , Queratinas/ultraestrutura , Camundongos , Peso Molecular , Nanopartículas/ultraestrutura , Oxirredução , Tamanho da Partícula , Ácido Peracético/química , Carneiro Doméstico , Engenharia Tecidual/métodos
14.
J Proteomics ; 186: 38-46, 2018 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-30030164

RESUMO

Mass spectrometry (MS) technology has a special advantage in species determination for protein-rich samples which requires identification of species-specific peptides. However, for species discrimination of highly homologous proteins, it remains challenging to select the species unique peptides with routine proteomics approaches. In this work, we chose keratins and keratin-associated proteins (KAPs) present in cashmere fibers from goat and wool fibers from sheep as targets, to develop a dual-protease digestion workflow based on in-silico and experimental analysis. Combined usage of Glu-C and trypsin proteases showed the best digestion performance for MS identification of keratins and KAPs from different species. The parallel reaction monitoring (PRM) technique was implemented to validate and quantify the selected species discriminable peptides. The fiber composition of both blended animal hair fibers and industrial textile fabrics were successfully determined with the PRM assay. Furthermore, we identified over 360 peptides from the cashmere fiber beyond the current Uniprot goat proteome database. We expect our new workflow would improve the identification and quantification of keratin and KAPs, and provide inspiration for distinguishing other highly homologous proteins. We also anticipate the set of species-specific peptides from keratin or KAPs validated in this work would benefit the quality assessment for industrial fiber materials and textile products. SIGNIFICANCE: Discriminating species from highly homologous proteins is challenging for MS-based shotgun proteomics. The large percentage of overlapped protein sequence hinders the identification of the species unique peptides. In this work, we aimed to discriminate sample species between goat and sheep from keratins and keratin-associated proteins (KAPs). A dedicated workflow was developed to boost the exposure and quantification of species discriminable peptides. The dual-proteases digestion approach was optimized based on amino acid sequence analysis and protein in-silico digestion analysis. The PRM assays were established to validate and quantify the selected species unique peptides. Additionally, we have identified about 360 novel candidate peptides complementary to the current goat protein sequence database. We expect our workflow would improve the species discrimination for highly homologous proteins and benefit the proteomics study of keratin and KAPs in the human proteome.


Assuntos
Queratinas/análise , Proteólise , Homologia de Sequência de Aminoácidos , Animais , Cabras , Cabelo/química , Espectrometria de Massas/métodos , Peptídeo Hidrolases/metabolismo , Peptídeos/análise , Especificidade da Espécie , Fibra de Lã/análise
15.
J Exp Biol ; 221(Pt 6)2018 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-29572427

RESUMO

Hair curvature underpins structural diversity and function in mammalian coats, but what causes curl in keratin hair fibres? To obtain structural data to determine one aspect of this question, we used confocal microscopy to provide in situ measurements of the two cell types that make up the cortex of merino wool fibres, which was chosen as a well-characterised model system representative of narrow diameter hairs, such as underhairs. We measured orthocortical and paracortical cross-sectional areas, and cortical cell lengths, within individual fibre snippets of defined uniplanar curvature. This allowed a direct test of two long-standing theories of the mechanism of curvature in hairs. We found evidence contradicting the theory that curvature results from there being more cells on the side of the fibre closest to the outside, or convex edge, of curvature. In all cases, the orthocortical cells close to the outside of curvature were longer than paracortical cells close to the inside of the curvature, which supports the theory that curvature is underpinned by differences in cell type length. However, the latter theory also implies that, for all fibres, curvature should correlate with the proportions of orthocortical and paracortical cells, and we found no evidence for this. In merino wool, it appears that the absolute length of cells of each type and proportion of cells varies from fibre to fibre, and only the difference between the length of the two cell types is important. Implications for curvature in higher diameter hairs, such as guard hairs and those on the human scalp, are discussed.


Assuntos
Queratinas/química , Fibra de Lã/análise , Lã/química , Animais , Contagem de Células , Carneiro Doméstico
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