PMID: 6403363Mar 1, 1983Paper

Purification and properties of a protein from bovine lens which inhibits trypsin and two endogenous lens proteinases

Experimental Eye Research
O P Srivastava, B J Ortwerth

Abstract

An inhibitor of trypsin-like proteinases was isolated from the water-soluble proteins of bovine lens cortex. The inhibitor was purified by four simple procedures: the separation of the inhibitor fraction by Agarose A-1.5 m gel filtration, extraction with 2.5% TCA at 70 degrees C, ammonium sulfate precipitation of the TCA-soluble proteins and a final separation by gel filtration chromatography. This preparation was found to be homogeneous by SDS-PAGE with an approximate subunit molecular weight of 5500 daltons. Gel filtration separated the ammonium sulfate precipitate into an inactive high-molecular-weight peak which eluted in the void volume, and two peaks of approximately 40 000 daltons and 10 000 daltons. Both low-molecular-weight peaks gave a single 5500 dalton band on SDS-PAGE, but only the 40 000 dalton peak was active when concentrated and assayed with bovine trypsin. These data suggest that the inhibitor is present in multimeric forms in solution, but only the octamer appears to be active. Antibodies prepared against the purified inhibitor showed a single precipitin line, while no reaction was seen with an alpha-crystallin antiserum. Upon storage in solution all of the inhibitor became converted into a high-molecular-wei...Continue Reading

References

Jun 1, 1964·The Biochemical Journal·S G Waley
May 1, 1981·Experimental Eye Research·S S Tse, B J Ortwerth
Jan 1, 1980·Annual Review of Biochemistry·M Laskowski, I Kato
Sep 1, 1980·Experimental Eye Research·S S Tse, B J Ortwerth

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Citations

Jan 1, 1989·Experimental Eye Research·O P Srivastava, B J Ortwerth
Dec 1, 1983·Experimental Eye Research·O P Srivastava, B J Ortwerth
May 1, 1983·Experimental Eye Research·O P Srivastava, B J Ortwerth
Sep 24, 1987·Biochimica Et Biophysica Acta·K K SharmaB J Ortwerth
Jun 1, 1992·Experimental Eye Research·O P SrivastavaK Srivastava

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