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PLoS One. 2015 Jul 14;10(7):e0132818. doi: 10.1371/journal.pone.0132818. eCollection 2015.

Design of a Selective Substrate and Activity Based Probe for Human Neutrophil Serine Protease 4.

Author information

1
Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland; Program in Cell Death and Survival Networks, Sanford-Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA 92024, United States of America.
2
Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
3
Program in Cell Death and Survival Networks, Sanford-Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA 92024, United States of America.
4
Dept. of Early Discovery Biochemistry, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States of America.

Abstract

Human neutrophil serine protease 4 (NSP4), also known as PRSS57, is a recently discovered fourth member of the neutrophil serine proteases family. Although its biological function is not precisely defined, it is suggested to regulate neutrophil response and innate immune reactions. To create optimal substrates and visualization probes for NSP4 that distinguish it from other NSPs we have employed a Hybrid Combinatorial Substrate Library approach that utilizes natural and unnatural amino acids to explore protease subsite preferences. Library results were validated by synthesizing individual substrates, leading to the identification of an optimal substrate peptide. This substrate was converted to a covalent diphenyl phosphonate probe with an embedded biotin tag. This probe demonstrated high inhibitory activity and stringent specificity and may be suitable for visualizing NSP4 in the background of other NSPs.

PMID:
26172376
PMCID:
PMC4501687
DOI:
10.1371/journal.pone.0132818
[Indexed for MEDLINE]
Free PMC Article

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