By Marie-Isabel Aguilar
Hands-on specialists from academia and comprehensively describe tips to effectively practice all of the serious HPLC strategies wanted for the research of peptides and proteins. The equipment diversity from normal suggestions to these for capillary to large-scale preparative isolation. The authors have additionally offered a few particular purposes as case reports to demonstrate the analytical methods to a selected separation or assay problem, with examples drawn from modern fields in biochemistry and biotechnology.
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Extra resources for HPLC of Peptides and Proteins: Methods and Protocols (Methods in Molecular Biology Vol 251)
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J Biol Chem 2002; 277:12190-12199. 97. Stein M P , Feng Y, Cooper KL et al. H u m a n VPS34 and p i 5 0 are Rab7 interacting partners. Traffic 2003; 4:754-771. 98. Ortiz D , Medkova M, Walch-Solimena C et al. Ypt32 recruits the Sec4p guanine nucleotide exchange factor, Sec2p, to secretory vesicles; evidence for a Rab cascade in yeast. J Cell Biol 2002; 157:1005-1015. 99. Walch-Solimena C, Collins RN, Novick P. Sec2 mediates nucleotide exchange on Sec4 and is involved in polarized delivery of post-Golgi vesicles.
Further studies of small GTPases and their effectors will be pivotal for our understanding of how membrane compartmentalization into domains may specify the identity and function of intracellular organelles. Acknowledgements The work in Marino Zerial s laboratory was supported by grants from the HFSP (RG-0260/ 1999-M), the European Union (HPRN-CT-2000-00081) and die Max Planck Society. References 1. McMahon HT, Mills IG. COP and clathrin-coated vesicle budding: Different pathways, common approaches.
Stenmark H , Parton RG, Steele-Mortimer O et al. Inhibition of rab5 GTPase activity stimulates membrane fusion in endocytosis. E M B O J 1994; 13:1287-1296. 39. Barbieri MA, Li G, Colombo M et al. Rab5, and early acting endosomal GTPase, supports in vitro endosome fusion without G T P hydrolysis. J Biol Chem 1994; 269:18720-18722. 40. Wucherpfennig T , Wilsch-Brauninger M, Gonzalez-Gaitan M . Role of Drosophila Rab5 during endosomal trafficking at the synapse and evoked neurotransmitter release.