A kinetic and equilibrium study of the denaturation of aspartic proteinases from the fungi, Endothia parasitica and Mucor miehei.


Abstract

Kinetic and equilibrium analyses of the denaturation of Endothia parasitica and Mucor miehei aspartic proteinases were performed using enzyme activity and ultraviolet absorption as indices of denaturation. Denaturation of these proteinases was shown to be irreversible, suggesting that the conformations of these aspartic proteinases may be predetermined in their zymogens. Thermal and guanidine hydrochloride denaturation of these proteinases produced first-order, two-state, kinetic behaviour. Equilibrium unfolding transitions of these proteinases were highly cooperative but not entirely coincident in the two indices employed, suggesting some deviation from two-state character. Oxidation to remove 37.8% of the carbohydrate of M. miehei glycoproteinase with sodium metaperiodate resulted in a substantial decrease in both kinetic and equilibrium stabilities without modification of the amino acid composition or specific activity. In addition, gel filtration subsequent to equilibrium studies indicated that partial removal of the carbohydrate from M. miehei proteinase promoted autolysis under denaturing conditions. Study holds ProTherm entries: 10030, 10031, 10032, 10033 Extra Details: aspartic proteinase; protein denaturation; kinetics; (E.parasitica);,(M.miehei)

Submission Details

ID: sWqqmFU

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:39 p.m.

Version: 1

Publication Details
Brown ED;Yada RY,Biochim. Biophys. Acta (1991) A kinetic and equilibrium study of the denaturation of aspartic proteinases from the fungi, Endothia parasitica and Mucor miehei. PMID:2001389
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2ASI 1996-08-01 2.15 ASPARTIC PROTEINASE
2RMP 1997-09-17 2.7 RMP-pepstatin A complex

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Mucorpepsin P00799 CARP_RHIMI