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

Debela, M., Magdolen, V., Skala, W., Elsasser, B., Schneider, E. L., Craik, C. S., et al. (2018). Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin. Scientific Reports, 8: 10705. doi:10.1038/s41598-018-29058-6.

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 Creators:
Debela, Mekdes1, Author              
Magdolen, Viktor2, Author
Skala, Wolfgang2, Author
Elsasser, Brigitta2, Author
Schneider, Eric L.2, Author
Craik, Charles S.2, Author
Biniossek, Martin L.2, Author
Schilling, Oliver2, Author
Bode, Wolfram1, Author              
Brandstetter, Hans2, Author
Goettig, Peter2, Author
Affiliations:
1Former Research Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565145              
2external, ou_persistent22              

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Free keywords: KALLIKREIN-RELATED PEPTIDASES; SERINE-PROTEASE INHIBITOR; NEUROPSIN MESSENGER-RNA; COAGULATION-FACTOR IXA; BINDING-SITE; CRYSTAL-STRUCTURES; MOUSE-BRAIN; TISSUE KALLIKREINS; BIOLOGICAL-FLUIDS; FACTOR XAScience & Technology - Other Topics;
 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.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published online
 Pages: 15
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
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Title: Scientific Reports
  Abbreviation : Sci. Rep.
Source Genre: Journal
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Publ. Info: London, UK : Nature Publishing Group
Pages: - Volume / Issue: 8 Sequence Number: 10705 Start / End Page: - Identifier: ISSN: 2045-2322
CoNE: https://pure.mpg.de/cone/journals/resource/2045-2322