Activity of human kallikrein-related peptidase 6 (KLK6) on substrates containing sequences of basic amino acids. Is it a processing protease?

Activity of human kallikrein-related peptidase 6 (KLK6) on substrates containing sequences of basic amino acids. Is it a processing protease?

Author Silva, Roberta N. Autor UNIFESP Google Scholar
Oliveira, Lilian C. G. Autor UNIFESP Google Scholar
Parise, Carolina B. Autor UNIFESP Google Scholar
Oliveira, Juliana R. Autor UNIFESP Google Scholar
Severino, Beatrice Google Scholar
Corvino, Angela Google Scholar
di Vaio, Paola Google Scholar
Temussi, Piero A. Google Scholar
Caliendo, Giuseppe Google Scholar
Santagada, Vincenzo Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Abstract Human kallikrein 6 (KLK6) is highly expressed in the central nervous system and with elevated level in demyelinating disease. KLK6 has a very restricted specificity for arginine (R) and hydrolyses myelin basic protein, protein activator receptors and human ionotropic glutamate receptor subunits. Here we report a previously unreported activity of KLK6 on peptides containing clusters of basic amino acids, as in synthetic fluorogenic peptidyl-Arg-7-amino-4-carbamoylmethylcoumarin (peptidyl-ACC) peptides and FRET peptides in the format of Abz-peptidyl-Q-EDDnp (where Abz = ortho-aminobenzoic acid and Q-EDDnp = glutaminyl-N-(2,4dinitrophenyl) ethylenediamine), in which pairs or sequences of basic amino acids (R or K) were introduced. Surprisingly, KLK6 hydrolyzed the fluorogenic peptides Bz-A-R down arrow R-ACC and Z-R down arrow R-MCA between the two R groups, resulting in non-fluorescent products. FRET peptides containing furin processing sequences of human MMP-14, nerve growth factor (NGF), Neurotrophin-3 (NT-3) and Neurotrophin-4 (NT-4) were cleaved by KLK6 at the same position expected by furin. Finally, KLK6 cleaved FRET peptides derived from human proenkephalin after the KR, the more frequent basic residues flanking enkephalins in human proenkephalin sequence. This result suggests the ability of KLK6 to release enkephalin from proenkephalin precursors and resembles furin a canonical processing proteolytic enzyme. Molecular models of peptides were built into the KLK6 structure and the marked preference of the cut between the two R of the examined peptides was related to the extended conformation of the substrates. (C) 2017 Elsevier B.V. All rights reserved.
Keywords Proteolytic enzymes
Plasma kallikrein
Tissue kallikrein
Matrix metalloproteases
xmlui.dri2xhtml.METS-1.0.item-coverage Amsterdam
Language English
Sponsor Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)
Instituto Nacional de Fluidos Complexos (INCT-FCx)
Grant number FAPESP: 12/50191-4R
CNPq: 471340/2011-1
CNPq: 470388/2010-2
INCT-FCx: 573560/2008-0
Date 2017
Published in Biochimica Et Biophysica Acta-Proteins And Proteomics. Amsterdam, v. 1865, n. 5, p. 558-564, 2017.
ISSN 1570-9639 (Sherpa/Romeo, impact factor)
Publisher Elsevier Science Bv
Extent 558-564
Access rights Closed access
Type Article
Web of Science ID WOS:000400714800011

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