Design and synthesis of new substrates of HtrA2 protease

Anal Biochem. 2015 Apr 15:475:44-52. doi: 10.1016/j.ab.2015.01.013. Epub 2015 Jan 29.

Abstract

HtrA2 belongs to the HtrA (high temperature requirement A) family of ATP-independent serine proteases. The primary function of HtrA2 includes maintaining the mitochondria homeostasis, cell death (by apoptosis, necrosis, or anoikis), and contribution to the cell signaling. Several recent reports have shown involvement of HtrA2 in development of cancer and neurodegenerative disorders. Here, we describe the profiling of HtrA2 protease substrate specificity via the combinatorial chemistry approach that led to the selection of novel intramolecularly quenched substrates. For all synthesized compounds, the highest HtrA2-mediated hydrolysis efficiency and selectivity among tested HtrA family members was observed for ABZ-Ile-Met-Thr-Abu-Tyr-Met-Phe-Tyr(3-NO2)-NH2, which displayed a specificity constant kcat/KM value of 14,535M(-1)s(-1).

Keywords: Fluorogenic substrates; HtrA2 proteinase; Peptide libraries; Proteolytic enzymes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • High-Temperature Requirement A Serine Peptidase 2
  • Humans
  • Mitochondrial Proteins / chemistry*
  • Peptides / chemical synthesis
  • Peptides / chemistry*
  • Serine Endopeptidases / chemistry*
  • Substrate Specificity

Substances

  • Mitochondrial Proteins
  • Peptides
  • Serine Endopeptidases
  • HTRA2 protein, human
  • High-Temperature Requirement A Serine Peptidase 2