Title
Protein arginine deiminase 4: evidence for a reverse protonation mechanism
UMMS Affiliation
Department of Biochemistry and Molecular Pharmacology
Publication Date
2007-06-05
Document Type
Article
Subjects
Amino Acid Sequence; Arginine; Binding Sites; Citrulline; Cysteine; Histidine; Humans; Hydrogen-Ion Concentration; Hydrolases; Kinetics; Molecular Sequence Data; Mutagenesis, Site-Directed; *Protein Processing, Post-Translational; *Protons; Recombinant Proteins; Substrate Specificity
Disciplines
Biochemistry | Enzymes and Coenzymes | Medicinal-Pharmaceutical Chemistry | Therapeutics
Abstract
The presumed role of an overactive protein arginine deiminase 4 (PAD4) in the pathophysiology of rheumatoid arthritis (RA) suggests that PAD4 inhibitors could be used to treat an underlying cause of RA, potentially offering a mechanism to stop further disease progression. Thus, the development of such inhibitors is of paramount importance. Toward the goal of developing such inhibitors, we initiated efforts to characterize the catalytic mechanism of PAD4 and thereby identify important mechanistic features that can be exploited for inhibitor development. Herein we report the results of mutagenesis studies as well as our efforts to characterize the initial steps of the PAD4 reaction, in particular, the protonation status of Cys645 and His471 prior to substrate binding. The results indicate that Cys645, the active site nucleophile, exists as the thiolate in the active form of the free enzyme. pH studies on PAD4 further suggest that this enzyme utilizes a reverse protonation mechanism.
DOI of Published Version
10.1021/bi700095s
Source
Biochemistry. 2007 Jun 5;46(22):6578-87. Epub 2007 May 12. Link to article on publisher's site
Journal/Book/Conference Title
Biochemistry
Related Resources
Repository Citation
Knuckley B, Bhatia M, Thompson PR. (2007). Protein arginine deiminase 4: evidence for a reverse protonation mechanism. Thompson Lab Publications. https://doi.org/10.1021/bi700095s. Retrieved from https://escholarship.umassmed.edu/thompson/74
Comments
At the time of publication, Paul Thompson was not yet affiliated with UMass Medical School.