Publication: Improved purification and fluorescence changes upon activation of human seminal plasma acidic protease proenzyme
Issued Date
1981-05-14
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ISSN
00052744
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2-s2.0-0019879693
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Mahidol University
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SCOPUS
Bibliographic Citation
BBA - Enzymology. Vol.659, No.1 (1981), 38-47
Suggested Citation
Piyawan Surinrut, Jisnuson Svasti, Rudee Surarit Improved purification and fluorescence changes upon activation of human seminal plasma acidic protease proenzyme. BBA - Enzymology. Vol.659, No.1 (1981), 38-47. doi:10.1016/0005-2744(81)90269-2 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/30228
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Title
Improved purification and fluorescence changes upon activation of human seminal plasma acidic protease proenzyme
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Abstract
Modifications have been made to the previous purification procedure so that electrophoretically homogeneous acidic protease (EC 3.4.23.-) proenzyme of specific activity 800 units/mg may be isolated from human seminal plasma with a yield of over 50%. The intrinsic fluorescence of the proenzyme shows maximum excitation and emission wavelengths at 280 and 340 nm, respectively, typical of proteins containing tryptophan. Complete activation causes a 30-35% decrease in intrinsic fluorescence, accompanied by a shift in λmaxto the blue of 4-6 nm. Time course studies indicate that acidification of proenzyme to pH 3.1 leads to a sudden large decrease in fluorescence that precedes both the appearance of active enzyme band on sodium dodecyl sulphate (SDS)-polyacrylamide gels and the generation of enzyme activity as detected by the turbidimetric milk clotting assay. These results suggest that acidification causes a rapid conformational change which promotes the release of the activation peptide from the proenzyme to yield the active enzyme. © 1981.