NDLI: Engineering trypsin for inhibitor resistance.

Please wait, while we are loading the content...

Year: 2017 Volume: 26

Year: 2016 Volume: 25

Year: 2015 Volume: 24

Year: 2015 Volume: 24 Issue: 12

Year: 2015 Volume: 24 Issue: 11

Year: 2015 Volume: 24 Issue: 10

Year: 2015 Volume: 24 Issue: 9

Membrane proteins in their native habitat as seen by solid-state NMR spectroscopy.

A unique binding mode of the eukaryotic translation initiation factor 4E for guiding the design of novel peptide inhibitors.

Peptide folding driven by Van der Waals interactions.

Residue-level global and local ensemble-ensemble comparisons of protein domains.

EsxB, a secreted protein from Bacillus anthracis forms two distinct helical bundles.

G protein-coupled odorant receptors: From sequence to structure.

Impact of cysteine variants on the structure, activity, and stability of recombinant human α-galactosidase A

Partners in crime: The role of tandem modules in gene transcription.

Construction of a robust and sensitive arginine biosensor through ancestral protein reconstruction.

Comparison of topological clustering within protein networks using edge metrics that evaluate full sequence, full structure, and active site microenvironment similarity.

Structural basis of the broadly neutralizing anti-interferon- antibody rontalizumab.

Structure-function correlations in tyrosinases.

Effect of single point mutations in a form of systemic amyloidosis.

Engineering trypsin for inhibitor resistance.

Comparison of intrinsic dynamics of cytochrome p450 proteins using normal mode analysis.

Structural mapping of the ClpB ATPases of Plasmodium falciparum: Targeting protein folding and secretion for antimalarial drug design.

Organization of the multiaminoacyl-tRNA synthetase complex and the cotranslational protein folding.

Linking structural features of protein complexes and biological function.

An ALS disease mutation in Cdc48/p97 impairs 20S proteasome binding and proteolytic communication.

Year: 2015 Volume: 24 Issue: 8

Year: 2015 Volume: 24 Issue: 7

Year: 2015 Volume: 24 Issue: 6

Year: 2015 Volume: 24 Issue: 5

Year: 2015 Volume: 24 Issue: 4

Year: 2015 Volume: 24 Issue: 3

Year: 2015 Volume: 24 Issue: 2

Year: 2015 Volume: 24 Issue: 1

Year: 2015 Volume: 24 Issue: Supplementary 1

Year: 2014 Volume: 23

Year: 2013 Volume: 22

Year: 2012 Volume: 21

Year: 2011 Volume: 20

Year: 2010 Volume: 19

Year: 2009 Volume: 18

Year: 2008 Volume: 17

Year: 2007 Volume: 16

Year: 2006 Volume: 15

Year: 2005 Volume: 14

Year: 2004 Volume: 13

Year: 2003 Volume: 12

Year: 2002 Volume: 11

Year: 2001 Volume: 10

Year: 2000 Volume: 9

Year: 1999 Volume: 8

Year: 1998 Volume: 7

Year: 1997 Volume: 6

Year: 1996 Volume: 5

Year: 1995 Volume: 4

Year: 1994 Volume: 3

Year: 1993 Volume: 2

Year: 1992 Volume: 1

Engineering trypsin for inhibitor resistance.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	St Germain, Commodore P. Gokey, Trevor Batt, Anna R. Guliaev, Anton B. Baird, Teaster
Description	Author Affiliation: Batt AR ( Department of Chemistry & Biochemistry, San Francisco State University, San Francisco, California, 94132.); St Germain CP ( Department of Chemistry & Biochemistry, San Francisco State University, San Francisco, California, 94132.); Gokey T ( Department of Chemistry & Biochemistry, San Francisco State University, San Francisco, California, 94132.); Guliaev AB ( Department of Chemistry & Biochemistry, San Francisco State University, San Francisco, California, 94132.); Baird T ( Department of Chemistry & Biochemistry, San Francisco State University, San Francisco, California, 94132.)
Abstract	The development of effective protease therapeutics requires that the proteases be more resistant to naturally occurring inhibitors while maintaining catalytic activity. A key step in developing inhibitor resistance is the identification of key residues in protease-inhibitor interaction. Given that majority of the protease therapeutics currently in use are trypsin-fold, trypsin itself serves as an ideal model for studying protease-inhibitor interaction. To test the importance of several trypsin-inhibitor interactions on the prime-side binding interface, we created four trypsin single variants Y39A, Y39F, K60A, and K60V and report biochemical sensitivity against bovine pancreatic trypsin inhibitor (BPTI) and M84R ecotin. All variants retained catalytic activity against small, commercially available peptide substrates [kcat /KM = (1.2 ± 0.3) × 10(7) M(-1 ) s(-1) . Compared with wild-type, the K60A and K60V variants showed increased sensitivity to BPTI but less sensitivity to ecotin. The Y39A variant was less sensitive to BPTI and ecotin while the Y39F variant was more sensitive to both. The relative binding free energies between BPTI complexes with WT, Y39F, and Y39A were calculated based on 3.5 µs combined explicit solvent molecular dynamics simulations. The BPTI:Y39F complex resulted in the lowest binding energy, while BPTI:Y39A resulted in the highest. Simulations of Y39F revealed increased conformational rearrangement of F39, which allowed formation of a new hydrogen bond between BPTI R17 and H40 of the variant. All together, these data suggest that positions 39 and 60 are key for inhibitor binding to trypsin, and likely more trypsin-fold proteases.
ISSN	09618368
e-ISSN	1469896X
DOI	10.1002/pro.2732
Journal	Protein Science
Issue Number	9
Volume Number	24
Language	English
Publisher	Wiley-Blackwell (on behalf of The Protein Society)
Publisher Date	2015-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Trypsin Inhibitors Chemistry Trypsin Amino Acid Sequence Animals Drug Resistance Kinetics Models, Molecular Molecular Dynamics Simulation Point Mutation Protein Binding Protein Conformation Protein Engineering Structure-activity Relationship Chemical Synthesis Trypsin Inhibitor, Kazal Pancreatic Pharmacology Research Support, N.i.h., Extramural Research Support, U.s. Gov't, Non-p.h.s. Discipline Biochemistry
Content Type	Text
Resource Type	Article
Subject	Medicine Molecular Biology Biochemistry

Central Library (ISO-9001:2015 Certified)
Indian Institute of Technology Kharagpur
Kharagpur, West Bengal, India | PIN - 721302

See location in the Map
03222 282435
Mail: support@ndl.gov.in

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in