Detection of Ras GTPase protein radicals through immuno-spin trapping. Free Radic Biol Med 2012 Sep 15;53(6):1339-45
Date
07/24/2012Pubmed ID
22819983Pubmed Central ID
PMC3549333DOI
10.1016/j.freeradbiomed.2012.07.009Scopus ID
2-s2.0-84865341861 (requires institutional sign-in at Scopus site) 10 CitationsAbstract
Over the past decade immuno-spin trapping (IST) has been used to detect and identify protein radical sites in numerous heme and metalloproteins. To date, however, the technique has had little application toward nonmetalloproteins. In this study, we demonstrate the successful application of IST in a system free of transition metals and present the first conclusive evidence of (•)NO-mediated protein radical formation in the HRas GTPase. HRas is a nonmetalloprotein that plays a critical role in regulating cell-growth control. Protein radical formation in Ras GTPases has long been suspected of initiating premature release of bound guanine nucleotide. This action results in altered Ras activity both in vitro and in vivo. As described herein, successful application of IST may provide a means for detecting and identifying radical-mediated Ras activation in many different cancers and disease states in which Ras GTPases play an important role.
Author List
Davis MF, Zhou L, Ehrenshaft M, Ranguelova K, Gunawardena HP, Chen X, Bonini MG, Mason RP, Campbell SLMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceAmino Acid Substitution
Cyclic N-Oxides
Diethylamines
Fourier Analysis
Free Radicals
Humans
Molecular Sequence Data
Molecular Weight
Nitric Oxide Donors
Oxidation-Reduction
Spin Labels
Spin Trapping
ras Proteins
