RESUMO
Enhancing factor (EF) protein, an isoform of secretory phospholipase A2 (PLA2), was purified as a modulator of epidermal growth factor from the small intestine of the Balb/c mouse. It was for the first time that a growth modulatory property of sPLA2 was demonstrated. Deletion mutation analysis of EF cDNA carried out in our laboratory showed that enhancing activity and phospholipase activity are two separate activities that reside in the same molecule. In order to study the specific amino acids involved in each of these activities, two site-directed mutants of EF were made and expressed in vitro. Comparison of enhancing activity as well as phospholipase A2 activity of these mutant proteins with that of wild type protein helped in identification of some of the residues important for both the activities
Assuntos
Animais , Sequência de Bases , Northern Blotting , Western Blotting , Linhagem Celular , Primers do DNA , Fosfolipases A2 do Grupo II , Humanos , Camundongos , Mutagênese Sítio-Dirigida , Fosfolipases A/genética , Fosfolipases A2RESUMO
Knowledge of candidate gene polymorphisms in a population is useful for a variety of gene-disease association studies, particularly for some complex traits. A number of candidate genes, a majority of them from the monoaminergic pathway in the brain, have been very popular in association studies with schizophrenia, a neuropsychiatric disorder. In this study diallelic/multiallelic polymorphisms in some dopaminergic, serotonergic and membrane-phospholipid-related genes have been evaluated in a control population recruited from North India. Association, if any, of these allelic variants with schizophrenia has been tested using a case-control approach. The case data have been taken from our published family-based association studies in schizophrenia. Of the eight genes tested in this study, association with schizophrenia was observed for only two gene polymorphisms, one in the promoter region of the serotonin 2A receptor gene and the other in the tryptophan hydroxylase gene. One new allele for the dopamine transporter gene (with eight repeats, 570-bp size), not reported in any population so far, has been identified in one individual in our sample. The data generated in this study, besides providing a normative background for various disease association studies, are a significant contribution to the population-specific genome database, a currently growing requirement.
Assuntos
Adulto , Estudos de Casos e Controles , Catecol O-Metiltransferase/genética , Dopamina/metabolismo , Feminino , Frequência do Gene , Humanos , Índia , Masculino , Fosfolipases A/genética , Monoéster Fosfórico Hidrolases/genética , Polimorfismo Genético , Receptores de Serotonina/genética , Esquizofrenia/genética , Triptofano Hidroxilase/genéticaRESUMO
Snake venom contains several toxins. Russell's viper (D. russellii, RV) is a venomous snake prevalent in northern and central Thailand. RV bites can cause disseminated coagulation, hemolysis, and edema of the bitten limbs. To identify protein components of RV venom, we made a cDNA library from RV venom glands, and randomly sequenced cloned cDNA. We were able to clone a cDNA encoding RV phospholipase A2 (PLA2). PLA2 is an active enzyme found in several species of snake venom worldwide. PLA2 is thought to be toxic to cell membrane, thereby, can cause local cell and tissue damage, as well as systemic effects in snake bite victims. This PLA2 cDNA clone would facilitate in vivo studies of the pathophysiology of RV bite.
Assuntos
Sequência de Aminoácidos , Animais , Clonagem Molecular , DNA Complementar/análise , Biblioteca Gênica , Fosfolipases A/genética , Fosfolipases A2 , Sensibilidade e Especificidade , Mordeduras de Serpentes/enzimologia , Tailândia/epidemiologia , Venenos de Víboras/enzimologia , ViperidaeRESUMO
Enhancing factor (EF) protein was initially purified as a modulator of epidermal growth factor from small intestines of mouse. The cDNA sequence, obtained by RT-PCR, revealed that EF belonged to the non-pancreatic, phospholipase A2 (PLA2) family. This was the first report of the mouse PLA2. In the present paper we report the complete cDNA sequence of EF gene, in which the 5' sequence has been obtained by RAcE-PCR. The predicted amino acid sequence was computer analysed and the putative sites for enzyme action, calcium binding and heparin binding have been identified. The complete protein sequence of EF along with 16 aligned sequences were used to infer a phylogenetic tree. From this data the mouse EF was grouped with other membrane associated PLA2 with a bootstrap value of 98% indicating that it belonged to this class.