Home Life Sciences Aspartate 496 from the subsite S2 drives specificity of human dipeptidyl peptidase III
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Aspartate 496 from the subsite S2 drives specificity of human dipeptidyl peptidase III

  • Marija Abramić EMAIL logo , Zrinka Karačić , Maja Šemanjski , Bojana Vukelić and Nina Jajčanin-Jozić
Published/Copyright: January 10, 2015

Abstract

Human dipeptidyl peptidase III (hDPP III) is a member of the M49 metallopeptidase family, which is involved in intracellular protein catabolism and oxidative stress response. To investigate the structural basis of hDPP III preference for diarginyl arylamide, using site-directed mutagenesis, we altered its S2 subsite to mimic the counterpart in yeast enzyme. Kinetic studies revealed that the single mutant D496G lost selectivity due to the increase of the Km value. The D496G, but not S504G, showed significantly decreased binding of peptides with N-terminal arginine, and of tynorphin. The results obtained identify Asp496 as an important determinant of human DPP III substrate specificity.


Corresponding author: Marija Abramić, Division of Organic Chemistry and Biochemistry, Ruder Bošković Institute, Bijenička cesta 54, P.O. Box 180, HR-10002 Zagreb, Croatia, e-mail:

Acknowledgments

Support for this study by the Ministry of Science, Education and Sport of the Republic of Croatia (project number 098-1191344-2938 to M.A.) and by the Alexander von Humboldt foundation (project name: ‘Study of plant enzymes from metallopeptidase families M20 and M49’) is gratefully acknowledged.

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Supplemental Material

The online version of this article (DOI: 10.1515/hsz-2014-0247) offers supplementary material, available to authorized users.


Received: 2014-9-9
Accepted: 2015-1-2
Published Online: 2015-1-10
Published in Print: 2015-4-1

©2015 by De Gruyter

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