A novel approach for screening the proteome for changes in protein conformation

Biochemistry
Anson PierceAsish Chaudhuri

Abstract

Changes in surface hydrophobicity are generally considered as a sensitive indicator for monitoring the structural alterations of proteins that are often associated with changes in function. Currently, no technique has been developed to screen a complex mixture of proteins for changes in the conformation of specific proteins. In this study, we adapted a UV photolabeling approach, using an apolar fluorescent probe, 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (BisANS), to monitor changes in surface hydrophobic domains in either purified rhodanese or skeletal muscle cytosolic proteins by urea-induced unfolding or in response to in vitro metal-catalyzed oxidation. Using two-dimensional polyacrylamide gel electrophoresis (2D PAGE), we identified two specific proteins in skeletal muscle cytosol that exhibited a marked loss of incorporation of BisANS after exposure to in vitro oxidative stress: creatine kinase (CK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found that the activities of both enzymes were also reduced significantly in response to oxidative stress. We then determined if this method could detect changes in surface hydrophobicity in specific proteins arising from oxidative stress generated in vivo by m...Continue Reading

References

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Citations

Feb 19, 2009·Proceedings of the National Academy of Sciences of the United States of America·Viviana I PérezAsish Chaudhuri
May 12, 2011·Integrative and Comparative Biology·Steven N Austad
Nov 21, 2013·Age·S B TreasterS N Austad
Feb 27, 2009·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Adam B SalmonAsish R Chaudhuri
Apr 12, 2007·Expert Review of Proteomics·Victor S Sharov, Christian Schöneich
Aug 18, 2009·Nitric Oxide : Biology and Chemistry·Lidiane Isabel FilippinRicardo Machado Xavier
Sep 1, 2015·Age·Heike GruberEva E R Philipp
Oct 24, 2006·Aging Cell·Blazej AndziakRochelle Buffenstein
Oct 1, 2008·Meat Science·M Gratacós-Cubarsí, R Lametsch
Apr 2, 2014·Biochemical and Biophysical Research Communications·Arunabh BhattacharyaAsish R Chaudhuri
Sep 5, 2014·PloS One·Rafael Medina-NavarroGenoveva Durán-Reyes
Jun 23, 2007·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Florian L MullerHolly Van Remmen
Nov 27, 2007·Free Radical Biology & Medicine·Anson P PierceAsish R Chaudhuri

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