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Methods Enzymol

Title:		"Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor"
Author(s):		Bader TK; Rappe TM; Veglia G; Distefano MD;
Address:		"University of Minnesota, Twin Cities, Minneapolis, MN, United States. University of Minnesota, Twin Cities, Minneapolis, MN, United States. Electronic address: diste001@umn.edu"
Journal Title:		Methods Enzymol
Year:		2019
Volume:		20181222
Issue:
Page Number:		207 - 238
DOI:		10.1016/bs.mie.2018.09.025
ISSN/ISBN:		1557-7988 (Electronic) 0076-6879 (Print) 0076-6879 (Linking)
Abstract:		"Protein and peptide prenylation is an essential biological process involved in many signal transduction pathways. Hence, it plays a critical role in establishing many major human ailments, including Alzheimer's disease, amyotrophic lateral sclerosis (ALS), malaria, and Ras-related cancers. Yeast mating pheromone a-factor is a small dodecameric peptide that undergoes prenylation and subsequent processing in a manner identical to larger proteins. Due to its small size in addition to its well-characterized behavior in yeast, a-factor is an attractive model system to study the prenylation pathway. Traditionally, chemical synthesis and characterization of a-factor have been challenging, which has limited its use in prenylation studies. In this chapter, a robust method for the synthesis of a-factor is presented along with a description of the characterization of the peptide using MALDI and NMR. Finally, complete assignments of resonances from the isoprenoid moiety and a-factor from COSY, TOCSY, HSQC, and long-range HMBC NMR spectra are presented. This methodology should be useful for the synthesis and characterization of other mature prenylated peptides and proteins"
Keywords:		"Chromatography, Affinity/methods Fluorenes/*chemistry Humans Mating Factor/chemical synthesis/*chemistry/isolation & purification Nuclear Magnetic Resonance, Biomolecular/*methods Peptides/chemical synthesis/*chemistry/isolation & purification Protein Pre;"
Notes:		"MedlineBader, Taysir K Rappe, Todd M Veglia, Gianlugi Distefano, Mark D eng R01 GM084152/GM/NIGMS NIH HHS/ R01 GM106082/GM/NIGMS NIH HHS/ RF1 AG056976/AG/NIA NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. 2019/01/07 Methods Enzymol. 2019; 614:207-238. doi: 10.1016/bs.mie.2018.09.025. Epub 2018 Dec 22"