Purification and light-dependent phosphorylation of a candidate fusion protein, the photoreceptor cell peripherin/rds

Biochemistry
K Boesze-BattagliaD S Williams

Abstract

The proteins peripherin/rds and rom-1 form a protein complex in the rims of photoreceptor outer segment disk membranes. Peripherin/rds plays an essential role in the morphogenesis and maintenance of disk membrane structure, with peripherin/rds gene mutations resulting in photoreceptor cell degeneration. We report two different chromatographic procedures for the purification of native peripherin/rds from bovine photoreceptor cell outer segments and show that the protein is a phosphoprotein that promotes membrane fusion in vitro. During one procedure, peripherin/rds was copurified in association with rom-1 by hyroxylapatite and Mono Q FPLC. During the other, it was purified free from rom-1 by concanavalin-A affinity chromatography and chromatofocusing. Analysis of homogeneous peripherin/rds from the second procedure showed that exposure of photoreceptor outer segments to light resulted in the incorporation of nearly 2 mol of phosphate per mole of peripherin/rds and a concomitant shift in the isoelectric point of the protein. In addition, we found that recombination of purified peripherin/rds into lipid vesicles increased membrane fusion, with more rapid fusion detected with phosphorylated peripherin/rds. In conclusion, studies wi...Continue Reading

Citations

Jun 10, 2011·Cellular and Molecular Life Sciences : CMLS·Shannon M ConleyMuna I Naash
Jun 1, 2005·Progress in Lipid Research·Arlene D Albert, Kathleen Boesze-Battaglia
Jun 17, 2000·Progress in Lipid Research·N M GiustoM G Ilincheta de Boschero
Jan 24, 2013·Investigative Ophthalmology & Visual Science·Françoise HaeseleerAmy Lee
Nov 16, 2007·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Amy LeeFrançoise Haeseleer
Jan 17, 2016·Progress in Retinal and Eye Research·Michael W StuckMuna I Naash
Jul 26, 2002·Experimental Eye Research·Kathleen Boesze-Battagliaa, Frank P Stefano
Jul 26, 2002·Experimental Eye Research·Susan Muller-WeeksCatherine Fitzgerald
Nov 14, 2006·International Review of Cytology·Andrew F X Goldberg
Oct 4, 2017·Stem Cell Research & Therapy·Roni A HazimDavid S Williams
Dec 9, 1998·Proceedings of the National Academy of Sciences of the United States of America·A E Otto-BrucK Palczewski
Apr 29, 1998·The Journal of Biological Chemistry·I P UdovichenkoD S Williams
Nov 8, 2013·The Journal of Biological Chemistry·Brian M KevanyKrzysztof Palczewski
Dec 14, 2004·The Journal of Biological Chemistry·Monika Damek-PoprawaKathleen Boesze-Battaglia
Oct 24, 2006·Experimental Eye Research·Kathleen Boesze-BattagliaSusan Muller-Weeks

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