Biochemical, immunological and kinetic characterization and partial sequence analysis of a thiol proteinase inhibitor from Bubalus bubalis kidney: An attempt targeting kidney disorders

Anas Shamsi; Azaj Ahmed; Bilqees Bano

doi:10.1016/j.ijbiomac.2015.12.084

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Biochemical, immunological and kinetic characterization and partial sequence analysis of a thiol proteinase inhibitor from Bubalus bubalis kidney: An attempt targeting kidney disorders

Journal article

Peer reviewed

Biochemical, immunological and kinetic characterization and partial sequence analysis of a thiol proteinase inhibitor from Bubalus bubalis kidney: An attempt targeting kidney disorders

Anas Shamsi, Azaj Ahmed and Bilqees Bano

International journal of biological macromolecules, Vol.94(Pt B), pp.819-826

01/2017

DOI: 10.1016/j.ijbiomac.2015.12.084

PMID: 26751400

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Abstract

In the present study a thiol proteinase inhibitor was isolated from buffalo kidney making use of ammonium sulphate precipitation and gel filtration chromatography on Sephacryl S-100HR column. Purified inhibitor is homogeneous as it displayed a single band in gel electrophoresis both under reducing and non-reducing environment and is of 65KDa as revealed by gel filtration and SDS PAGE. Kinetic studies revealed the presence of reversible accompanied with competitive mode of inhibition; showing maximum efficacy against papain (K =2.90×10 ). It was maximally active at pH 8.0 and was stable for a period of 30, 60 and 90 days at 37, 4 and -20°C respectively. Immunological studies confirmed its purity of epitopes as a single precipitin line is obtained in immunodiffusion. N-terminal analysis revealed that it shared a good homology with mouse kidney cystatin as well as with Human Cys C and Cys E thereby advocating its use as a model for various human oriented studies which targets how the kidney cystatin level varies in accordance with various drugs that are currently being used as a target for variety of diseases.

Amino Acid Sequence

Animals

Bromelains - antagonists & inhibitors

Bromelains - chemistry

Buffaloes

Cystatins - chemistry

Cystatins - immunology

Cystatins - isolation & purification

Ficain - antagonists & inhibitors

Ficain - chemistry

Humans

Hydrogen-Ion Concentration

Kidney - chemistry

Kidney - immunology

Kinetics

Mice

Molecular Weight

Papain - antagonists & inhibitors

Papain - chemistry

Protease Inhibitors - chemistry

Protease Inhibitors - immunology

Protease Inhibitors - isolation & purification

Protein Stability

Sequence Alignment

Sulfhydryl Compounds - chemistry

Details

Title: Subtitle: Biochemical, immunological and kinetic characterization and partial sequence analysis of a thiol proteinase inhibitor from Bubalus bubalis kidney: An attempt targeting kidney disorders
Creators: Anas Shamsi - Aligarh Muslim University
Azaj Ahmed - Aligarh Muslim University
Bilqees Bano - Aligarh Muslim University
Resource Type: Journal article
Publication Details: International journal of biological macromolecules, Vol.94(Pt B), pp.819-826
DOI: 10.1016/j.ijbiomac.2015.12.084
PMID: 26751400
NLM abbreviation: Int J Biol Macromol
ISSN: 0141-8130
eISSN: 1879-0003
Language: English
Date published: 01/2017
Academic Unit: Internal Medicine
Record Identifier: 9985177941502771

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