title: Design of a Selective Substrate and Activity Based Probe for Human Neutrophil Serine Protease 4.
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.
authors.affiliation: 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.
authors.forename: Paulina
authors.initials: P
authors.lastname: Kasperkiewicz
authors.affiliation: Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Poreba
authors.affiliation: 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.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Snipas
authors.affiliation: Dept. of Early Discovery Biochemistry, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States of America.
authors.forename: S Jack
authors.initials: SJ
authors.lastname: Lin
authors.affiliation: Dept. of Early Discovery Biochemistry, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States of America.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kirchhofer
authors.affiliation: 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.
authors.forename: Guy S
authors.initials: GS
authors.lastname: Salvesen
authors.affiliation: Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Drag
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D057057:Serine Proteases / Q000378:metabolism*; D013379:Substrate Specificity / Q000276:immunology* / Q000502:physiology
chemical_list: D000596:Amino Acids; D010455:Peptides; D057057:Serine Proteases