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.
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.
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Articles in the same Issue
- Frontmatter
- Reviews
- Sortase-mediated backbone cyclization of proteins and peptides
- Making the LINC: SUN and KASH protein interactions
- Enhancers, enhancers – from their discovery to today’s universe of transcription enhancers
- Minireview
- Recent advances and concepts in substrate specificity determination of proteases using tailored libraries of fluorogenic substrates with unnatural amino acids
- Research Articles/Short Communications
- Genes and Nucleic Acids
- microRNA-210 is involved in the regulation of postmenopausal osteoporosis through promotion of VEGF expression and osteoblast differentiation
- Protein Structure and Function
- C-terminal truncation of a Tat passenger protein affects its membrane translocation by interfering with receptor binding
- Aspartate 496 from the subsite S2 drives specificity of human dipeptidyl peptidase III
- Proteolysis
- Evolutionary divergence of Threonine Aspartase1 leads to species-specific substrate recognition
- Purification and characterisation of recombinant His-tagged RgpB gingipain from Porphymonas gingivalis