Structural characterization of human glyoxalase II as probed by limited proteolysis

Human glyoxalase II is partially proteolyzed by trypsin, under non denaturing conditions, only at the level of the C‐terminal region. The proteolytic cleavage resulted in an inactivation of the enzyme without loss of the secondary structure. Sodium dodecyl sulphate polyacrylamide gel‐electrophoresis...

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Bibliographic Details
Published in:Biochemistry and molecular biology international 1998-04, Vol.44 (4), p.761-769
Main Authors: Aceto, Antonio, Dragani, Beatrice, Melino, Sonia, Petruzzelli, Raffaele, Gualtieri, Gabriella, Principato, Giovanni, Saccucci, Franca
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Binding Sites
Circular Dichroism
domains
Enzyme Activation
Glyoxalase II
Humans
limited proteolysis
Molecular Sequence Data
Protein Folding
Protein Structure, Secondary
Thiolester Hydrolases - metabolism
Trypsin - metabolism
Trypsin - pharmacology
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Summary:Human glyoxalase II is partially proteolyzed by trypsin, under non denaturing conditions, only at the level of the C‐terminal region. The proteolytic cleavage resulted in an inactivation of the enzyme without loss of the secondary structure. Sodium dodecyl sulphate polyacrylamide gel‐electrophoresis and microsequence analysis showed that the glyoxalase II is proteolyzed at the level of Arg 184 and Lys 230 and undergoes a third cleavage in a region located at the beginning of the supposed C‐terminal domain. The proteolysis occurs either in the presence or in the absence of specific inhibitors. Our limited proteolysis experiments and secondary structure prediction give evidence for the presence of two domains characterized by different pattern of secondary structure.
ISSN:1521-6543
1039-9712
1521-6551
DOI:10.1080/15216549800201802