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Purification, characterization and kinetics of thiol protease inhibitor from goat (Capra hircus) lung

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Abstract

In the present study, two molecular forms of goat lung cystatin (GLC), I and II, were purified to homogeneity by a two-step procedure including ammonium sulfate precipitation (40–60%) and ion exchange chromatography. The inhibitor forms migrated as single bands under native and SDS-PAGE with and without reducing agent giving molecular mass of 66.4 and 76.4 kDa, respectively. GLC-I possesses 0.07% and GLC-II 2.3% carbohydrate content and no -SH groups. GLC-I showed greater affinity for papain than for ficin and bromelain. Immunological studies showed that the inhibitor was pure and there was cross reactivity between anti-GLC-I serum and goat brain cystatin. Both inhibitor forms were stable in the pH range of 3–10 and up to 75°C. GLC-I was found to possess 49% α-helical structure by CD spectroscopy. The inhibitor-papain complexes showed conformational changes as invoked by UV and fluorescence spectroscopic studies.

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Abbreviations

GLC:

goat lung cystatin

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, India

    Mohd Shahnawaz Khan & Bilqees Bano

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  1. Mohd Shahnawaz Khan
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  2. Bilqees Bano
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Correspondence to Bilqees Bano.

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Published in Russian in Biokhimiya, 2009, Vol. 74, No. 7, pp. 963–971.

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Khan, M.S., Bano, B. Purification, characterization and kinetics of thiol protease inhibitor from goat (Capra hircus) lung. Biochemistry Moscow 74, 781–788 (2009). https://doi.org/10.1134/S0006297909070116

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