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Isolation, characterization and kinetics of goat cystatins.
Comp Biochem Physiol B Biochem Mol Biol. 2005 Dec; 142(4):361-8.CB

Abstract

Two cysteine proteinase inhibitors I and II were purified from goat kidney using alkaline denaturation, ammonium sulphate fractionation, gel filtration on Sephadex G-75 and ion exchange chromatography on DEAE cellulose. The purified inhibitors were homogenous and showed a single band on SDS PAGE under reducing and non-reducing conditions with an apparent molecular mass of 67 kDa. The cystatin forms were stable in the range of pH 3-10 and up to 95 degrees C. Immunological identity with the sheep LMW kininogen was obtained suggesting that the inhibitor is closely related to kininogens. Spectral studies confirm that the inhibitors have predominantly an alpha-helical structure and undergo major conformational changes during complex formation with papain. The inhibitors had similar inhibitory activities on cysteine proteinases. Both inhibitors inhibited papain, ficin and bromelain competitively, with maximum affinity for papain. The overall lower affinity of these inhibitors to cysteine proteinases compared to other known cystatins can be attributed to the unusual N-terminal sequence where Leu is substituted by Ile. Furthermore, N-terminal sequence analysis revealed maximum homology to mammalian LMW kininogen.

Authors+Show Affiliations

Sadaf Z
Department of Biochemistry, F/O of Life Science, Aligarh Muslim University, UP, India. sadafzk@yahoo.co.in
Shahid PB
No affiliation info available
Bilqees B
No affiliation info available

MeSH

Amino Acid SequenceAnimalsAnti-Bacterial AgentsChromatography, Ion ExchangeCystatinsCysteine Proteinase InhibitorsElectrophoresis, Polyacrylamide GelGoatsHydrogen-Ion ConcentrationKidneyKineticsMolecular Sequence DataMolecular WeightPapainProtein BindingSpecies Specificity

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16257555

Citation

Sadaf, Z, et al. "Isolation, Characterization and Kinetics of Goat Cystatins." Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology, vol. 142, no. 4, 2005, pp. 361-8.
Sadaf Z, Shahid PB, Bilqees B. Isolation, characterization and kinetics of goat cystatins. Comp Biochem Physiol B Biochem Mol Biol. 2005;142(4):361-8.
Sadaf, Z., Shahid, P. B., & Bilqees, B. (2005). Isolation, characterization and kinetics of goat cystatins. Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology, 142(4), 361-8.
Sadaf Z, Shahid PB, Bilqees B. Isolation, Characterization and Kinetics of Goat Cystatins. Comp Biochem Physiol B Biochem Mol Biol. 2005;142(4):361-8. PubMed PMID: 16257555.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation, characterization and kinetics of goat cystatins. AU - Sadaf,Z, AU - Shahid,P B, AU - Bilqees,B, Y1 - 2005/10/28/ PY - 2004/04/02/received PY - 2005/08/18/revised PY - 2005/08/21/accepted PY - 2005/11/1/pubmed PY - 2006/3/10/medline PY - 2005/11/1/entrez SP - 361 EP - 8 JF - Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology JO - Comp Biochem Physiol B Biochem Mol Biol VL - 142 IS - 4 N2 - Two cysteine proteinase inhibitors I and II were purified from goat kidney using alkaline denaturation, ammonium sulphate fractionation, gel filtration on Sephadex G-75 and ion exchange chromatography on DEAE cellulose. The purified inhibitors were homogenous and showed a single band on SDS PAGE under reducing and non-reducing conditions with an apparent molecular mass of 67 kDa. The cystatin forms were stable in the range of pH 3-10 and up to 95 degrees C. Immunological identity with the sheep LMW kininogen was obtained suggesting that the inhibitor is closely related to kininogens. Spectral studies confirm that the inhibitors have predominantly an alpha-helical structure and undergo major conformational changes during complex formation with papain. The inhibitors had similar inhibitory activities on cysteine proteinases. Both inhibitors inhibited papain, ficin and bromelain competitively, with maximum affinity for papain. The overall lower affinity of these inhibitors to cysteine proteinases compared to other known cystatins can be attributed to the unusual N-terminal sequence where Leu is substituted by Ile. Furthermore, N-terminal sequence analysis revealed maximum homology to mammalian LMW kininogen. SN - 1096-4959 UR - https://www.unboundmedicine.com/medline/citation/16257555/Isolation_characterization_and_kinetics_of_goat_cystatins_ DB - PRIME DP - Unbound Medicine ER -