Peptide Microarrays for the Determination of Protease Substrate Specificity

2002 ◽  
Vol 124 (50) ◽  
pp. 14868-14870 ◽  
Author(s):  
Cleo M. Salisbury ◽  
Dustin J. Maly ◽  
Jonathan A. Ellman
Keyword(s):  
Substrate Specificity ◽  
Peptide Microarrays ◽  
Protease Substrate

Profiling serine protease substrate specificity with solution phase fluorogenic peptide microarrays

PROTEOMICS ◽  
2005 ◽  
Vol 5 (5) ◽  
pp. 1292-1298 ◽  
Author(s):  
Dhaval N. Gosalia ◽  
Cleo M. Salisbury ◽  
Dustin J. Maly ◽  
Jonathan A. Ellman ◽  
Scott L. Diamond
Keyword(s):  
Substrate Specificity ◽  
Serine Protease ◽  
Solution Phase ◽  
Peptide Microarrays ◽  
Protease Substrate ◽  
Fluorogenic Peptide

scholarly journals Rapid Determination of Substrate Specificity ofClostridium histolyticumβ-Collagenase Using an Immobilized Peptide Library

2001 ◽  
Vol 277 (10) ◽  
pp. 8366-8371 ◽  
Author(s):  
Yongbo Hu ◽  
Erin Webb ◽  
Jasbir Singh ◽  
Barry A. Morgan ◽  
James A. Gainor ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Rapid Determination ◽  
Peptide Library ◽  
Immobilized Peptide

scholarly journals Determination of extended substrate specificity of the MALT1 as a strategy for the design of potent substrates and activity-based probes

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Paulina Kasperkiewicz ◽  
Sonia Kołt ◽  
Tomasz Janiszewski ◽  
Katarzyna Groborz ◽  
Marcin Poręba ◽  
...  
Keyword(s):  
Substrate Specificity

scholarly journals ZapA, a possible virulence factor from Proteus mirabilis exhibits broad protease substrate specificity

2001 ◽  
Vol 34 (11) ◽  
pp. 1397-1403 ◽  
Author(s):  
M.A.F. Anéas ◽  
F.C.V. Portaro ◽  
I. Lebrun ◽  
L. Juliano ◽  
M.S. Palma ◽  
...  
Keyword(s):  
Substrate Specificity ◽  
Virulence Factor ◽  
Proteus Mirabilis ◽  
Protease Substrate

scholarly journals Structure determination of Murine Norovirus NS6 proteases with C-terminal extensions designed to probe protease-substrate interactions

2014 ◽  
Author(s):  
Humberto Fernandes ◽  
Eoin N Leen ◽  
Hamlet Cromwell Jnr ◽  
Marc-Philipp Pfeil ◽  
Stephen Curry
Keyword(s):  
Structure Determination ◽  
Crystal Form ◽  
Molecular Replacement ◽  
Murine Norovirus ◽  
Substrate Interactions ◽  
C Terminus ◽  
Peptide Binding Site ◽  
Specific Protease ◽  
Protease Substrate

Noroviruses are positive-sense single-stranded RNA viruses. They encode an NS6 protease that cleaves a viral polyprotein at specific sites to produce mature viral proteins. In an earlier study we obtained crystals of murine norovirus (MNV) NS6 protease in which crystal contacts were mediated by specific insertion of the C-terminus of one protein (which contains residues P5-P1 of the NS6-7 cleavage junction) into the peptide binding site of an adjacent molecule, forming an adventitious protease-product complex. We sought to reproduce this crystal form to investigate protease-substrate complexes by extending the C-terminus of NS6 construct to include residues on the C-terminal (P´) side of the cleavage junction. We report the crystallization and crystal structure determination of inactive mutants of murine norovirus NS6 protease with C-terminal extensions of one, two and four residues from the N-terminus of the adjacent NS7 protein (NS6 1´, NS6 2´, NS6 4´). We also determined the structure of a chimeric extended NS6 protease in which the P4 P4′ sequence of the NS6-7 cleavage site was replaced with the corresponding sequence from the NS2-3 cleavage junction (NS6 4´ 2|3). The constructs NS6 1′ and NS6 2′ yielded crystals that diffracted anisotropically. We found that, although the uncorrected data could be phased by molecular replacement, refinement of the structures stalled unless the data were ellipsoidally truncated and corrected with anisotropic B-factors. These corrections significantly improved phasing by molecular replacement and subsequent refinement. The refined structures of all four extended NS6 proteases are very similar in structure to the mature MNV NS6 — and in one case reveal additional details of a surface loop. Although the packing arrangement observed showed some similarities to those observed in the adventitious protease-product crystals reported previously, in no case were specific protease-substrate interactions observed.

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