PMID: 9659394Jul 11, 1998Paper

A study of the specificity of barley chymotrypsin inhibitor 2 by cysteine engineering of the P1 residue

Biochimica Et Biophysica Acta
Z Hasan, R J Leatherbarrow

Abstract

A combination of oligonucleotide-directed mutagenesis and chemical modification was used to produce reactive site (P1) variants of chymotrypsin inhibitor II (CI2) in an attempt to create more potent inhibitors and examine inhibitory specificity. Mutagenesis to introduce a unique cysteine (CI2M59C) followed by modification to S-carboxamidocysteine with iodoacetamide produced a 7-fold more potent inhibitor of subtilisin BPN' than the wild type inhibitor. Modification with iodoacetic acid, which gives a negatively charged P1 residue (S-carboxymethylcysteine), generates a weaker inhibitor of subtilisin BPN' and chymotrypsin. Further chemical modification experiments of CI2M59C with a series of iodoalkanes of increasing chain lengths was used to determine the optimal P1 side chain length required for potent inhibition of porcine pancreatic elastase. A trend was observed which implies that for CI2 the original methionine residue or its isostere S-ethylcysteine are most effective residues at this position and not alanine as might have been expected from the substrate specificity. The mutant CI2M59C did not inhibit human neutrophil elastase. The iodoalkane modifications not only resulted in recovery of inhibitory activity but also prov...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Nov 1, 1979·Analytical Biochemistry·E G DelMarM C Geokas
Dec 1, 1975·Biochemical Pharmacology·S Cha
Feb 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
Jan 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·T A Kunkel
Jul 17, 1973·Biochemical and Biophysical Research Communications·J Bieth, C G Wermuth
Apr 20, 1967·Biochemical and Biophysical Research Communications·I Schechter, A Berger
Jun 30, 1994·Biochimica Et Biophysica Acta·M A GreaggR J Leatherbarrow
May 1, 1985·Physical Review. B, Condensed Matter·M Schreiber
May 1, 1959·Archives of Biochemistry and Biophysics·G L ELLMAN
Nov 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·C A McPhalenM N James
Dec 1, 1987·Journal of Religion and Health·J W Jones

❮ Previous
Next ❯

Citations

Nov 26, 2010·Nucleic Acids Research·Michael T MorganAlexander C Drohat
Jan 18, 2014·Nucleic Acids Research·Megan E FitzgeraldAnna Marie Pyle
Aug 3, 2016·FEBS Letters·Demetra S M ChatzileontiadouDemetres D Leonidas
Feb 18, 2009·The FEBS Journal·Georgia A Kotzia, Nikolaos E Labrou

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.