Analysis of small GTP-binding proteins of the lens by GTP overlay assay reveals the presence of unique GTP-binding proteins associated with fiber cells

Experimental Eye Research
P V RaoD Garland

Abstract

Low molecular weight GTP-binding proteins are molecular switches which are thought to play pivotal roles in cell growth, differentiation, cytoskeletal organization and vesicular trafficking. In this study, members of this family of proteins have been identified and characterized in the eye lens, for the first time. [alpha 33P]GTP blot overlay assays of monkey and human lens water soluble and membranous insoluble fractions revealed the presence of specific GTP-binding proteins in the range of 20-30 kDa (small GTPases) in both fractions, with much higher amounts in the membranous insoluble fraction. In the insoluble fraction, in addition to 20-30 kDa GTPases, there are three distinct GTP-binding proteins, ranging from 33-45 kDa. The small GTPases (20-30 kDa) were present throughout the lens in epithelium, cortex and nucleus, while the 33-45 kDa GTP-binding protein bands were exclusively associated with the cortex and nucleus (fiber cells). Analysis of lens fractions by two-dimensional electrophoresis, immunoprecipitation using monoclonal and sequence specific polyclonal antibodies and C3 exoenzyme mediated ADP-ribosylation demonstrated the presence of Ras, Rap, Rho, Rac, Rab and several other small GTPases. The 33-45 kDa GTP-bind...Continue Reading

Citations

Jan 4, 2007·Assay and Drug Development Technologies·Maria E Verdugo-GazdikMark W Tengowski
Jan 11, 2007·Seminars in Cell & Developmental Biology·Y ChenJ W McAvoy
Dec 6, 2006·Seminars in Cell & Developmental Biology·P Vasantha Rao, Rupalatha Maddala
Jul 12, 2001·Experimental Eye Research·Q F ChengJ S Zigler
Apr 27, 2012·Microbiological Research·Kannan PalaniyandiSujatha Narayanan
Sep 28, 2017·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Lianna R WalkerDaniel C Ciobanu
Feb 19, 2002·Laboratory Investigation; a Journal of Technical Methods and Pathology·Vasantha RaoSamuel Zigler

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