Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin

Abstract Human KLK8/neuropsin, a kallikrein-related serine peptidase, is mostly expressed in skin and the hippocampus regions of the brain, where it regulates memory formation by synaptic remodeling. Substrate profiles of recombinant KLK8 were analyzed with positional scanning using fluorogenic tetr...

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Autores principales:	Mekdes Debela, Viktor Magdolen, Wolfgang Skala, Brigitta Elsässer, Eric L. Schneider, Charles S. Craik, Martin L. Biniossek, Oliver Schilling, Wolfram Bode, Hans Brandstetter, Peter Goettig
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c16e2c7087564dea9f129dcd30f35722
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spelling	oai:doaj.org-article:c16e2c7087564dea9f129dcd30f357222021-12-02T11:40:25ZStructural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin10.1038/s41598-018-29058-62045-2322https://doaj.org/article/c16e2c7087564dea9f129dcd30f357222018-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-29058-6https://doaj.org/toc/2045-2322Abstract Human KLK8/neuropsin, a kallikrein-related serine peptidase, is mostly expressed in skin and the hippocampus regions of the brain, where it regulates memory formation by synaptic remodeling. Substrate profiles of recombinant KLK8 were analyzed with positional scanning using fluorogenic tetrapeptides and the proteomic PICS approach, which revealed the prime side specificity. Enzyme kinetics with optimized substrates showed stimulation by Ca2+ and inhibition by Zn2+, which are physiological regulators. Crystal structures of KLK8 with a ligand-free active site and with the inhibitor leupeptin explain the subsite specificity and display Ca2+ bound to the 75-loop. The variants D70K and H99A confirmed the antagonistic role of the cation binding sites. Molecular docking and dynamics calculations provided insights in substrate binding and the dual regulation of activity by Ca2+ and Zn2+, which are important in neuron and skin physiology. Both cations participate in the allosteric surface loop network present in related serine proteases. A comparison of the positional scanning data with substrates from brain suggests an adaptive recognition by KLK8, based on the tertiary structures of its targets. These combined findings provide a comprehensive picture of the molecular mechanisms underlying the enzyme activity of KLK8.Mekdes DebelaViktor MagdolenWolfgang SkalaBrigitta ElsässerEric L. SchneiderCharles S. CraikMartin L. BiniossekOliver SchillingWolfram BodeHans BrandstetterPeter GoettigNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-15 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mekdes Debela Viktor Magdolen Wolfgang Skala Brigitta Elsässer Eric L. Schneider Charles S. Craik Martin L. Biniossek Oliver Schilling Wolfram Bode Hans Brandstetter Peter Goettig Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
description	Abstract Human KLK8/neuropsin, a kallikrein-related serine peptidase, is mostly expressed in skin and the hippocampus regions of the brain, where it regulates memory formation by synaptic remodeling. Substrate profiles of recombinant KLK8 were analyzed with positional scanning using fluorogenic tetrapeptides and the proteomic PICS approach, which revealed the prime side specificity. Enzyme kinetics with optimized substrates showed stimulation by Ca2+ and inhibition by Zn2+, which are physiological regulators. Crystal structures of KLK8 with a ligand-free active site and with the inhibitor leupeptin explain the subsite specificity and display Ca2+ bound to the 75-loop. The variants D70K and H99A confirmed the antagonistic role of the cation binding sites. Molecular docking and dynamics calculations provided insights in substrate binding and the dual regulation of activity by Ca2+ and Zn2+, which are important in neuron and skin physiology. Both cations participate in the allosteric surface loop network present in related serine proteases. A comparison of the positional scanning data with substrates from brain suggests an adaptive recognition by KLK8, based on the tertiary structures of its targets. These combined findings provide a comprehensive picture of the molecular mechanisms underlying the enzyme activity of KLK8.
format	article
author	Mekdes Debela Viktor Magdolen Wolfgang Skala Brigitta Elsässer Eric L. Schneider Charles S. Craik Martin L. Biniossek Oliver Schilling Wolfram Bode Hans Brandstetter Peter Goettig
author_facet	Mekdes Debela Viktor Magdolen Wolfgang Skala Brigitta Elsässer Eric L. Schneider Charles S. Craik Martin L. Biniossek Oliver Schilling Wolfram Bode Hans Brandstetter Peter Goettig
author_sort	Mekdes Debela
title	Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
title_short	Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
title_full	Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
title_fullStr	Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
title_full_unstemmed	Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin
title_sort	structural determinants of specificity and regulation of activity in the allosteric loop network of human klk8/neuropsin
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/c16e2c7087564dea9f129dcd30f35722
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_version_	1718395668558512128

Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin

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