PXD006884

PXD006884 is an original dataset announced via ProteomeXchange.

Title	Structural determinants of specificity and antagonistic Ca2+ and Zn2+ regulation of peptidase activity in human KLK8/neuropsin
Description	Human KLK8/neuropsin, a kallikrein-related serine peptidase, is mostly expressed in skin and hippocampus, where it regulates memory formation by synaptic remodeling. Recombinantly expressed KLK8 revealed a good accordance two substrate profiling methods: positional scanning with fluorogenic tetrapeptides and a novel version of the proteomic PICS approach. Besides a strong preference for Arg in the P1 position, the enzyme favors Thr in P4, basic P3 residues, aliphatic P2, small P1′ and aliphatic P2′ residues. Enzyme kinetic studies with synthesized substrates showed stimulatory and inhibitory effects of Ca2+ and Zn2+, respectively, which are important for physiological functions. Two crystal structures of KLK8 with the active site inhibitor leupeptin explain the subsite specificity and display Ca2+ bound to the 75-loop, which is unique among the KLKs. The mutants D70K and H99A confirmed the antagonistic role of the cation binding sites, for which a comparison with the murine apo-KLK8 structure clarified further mechanistic details. Molecular docking and dynamics calculations provided insights in substrate binding and the dual regulation by Ca2+ and Zn2+, involving an allosteric surface loop network.
HostingRepository	PRIDE
AnnounceDate	2018-10-17
AnnouncementXML	Submission_2018-10-17_01:40:33.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Oliver Schilling
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-07-06 05:14:22	ID requested
⏵ 1	2018-10-17 01:40:34	announced

Debela M, Magdolen V, Skala W, Els, ä, sser B, Schneider EL, Craik CS, Biniossek ML, Schilling O, Bode W, Brandstetter H, Goettig P, Structural determinants of specificity and regulation of activity in the allosteric loop network of human KLK8/neuropsin. Sci Rep, 8(1):10705(2018) [pubmed]

curator keyword: Biological

submitter keyword: Protease, peptide library

Oliver Schilling
contact affiliation	nstitute of Molecular Medicine and Cell Research University of Freiburg Stefan-Meier-Str. 17, Room 02 027 D-79104 Freiburg, Germany +49 761 203 9615 oliver.schilling@mol-med.uni-freiburg.de
contact email	oliver.schilling@mol-med.uni-freiburg.de
lab head
Oliver Schilling
contact affiliation	University of Freiburg
contact email	oliver.schilling@mol-med.uni-freiburg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD006884
     1. Label: PRIDE project
     2. Name: Structural determinants of specificity and antagonistic Ca2+ and Zn2+ regulation of peptidase activity in human KLK8/neuropsin