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Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor.
- Source :
-
Methods in enzymology [Methods Enzymol] 2019; Vol. 614, pp. 207-238. Date of Electronic Publication: 2018 Dec 22. - Publication Year :
- 2019
-
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.<br /> (© 2019 Elsevier Inc. All rights reserved.)
- Subjects :
- Chromatography, Affinity methods
Humans
Mating Factor chemical synthesis
Mating Factor isolation & purification
Peptides chemical synthesis
Peptides isolation & purification
Protein Prenylation
Saccharomyces cerevisiae metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
Trityl Compounds chemistry
Fluorenes chemistry
Mating Factor chemistry
Nuclear Magnetic Resonance, Biomolecular methods
Peptides chemistry
Saccharomyces cerevisiae chemistry
Solid-Phase Synthesis Techniques methods
Subjects
Details
- Language :
- English
- ISSN :
- 1557-7988
- Volume :
- 614
- Database :
- MEDLINE
- Journal :
- Methods in enzymology
- Publication Type :
- Academic Journal
- Accession number :
- 30611425
- Full Text :
- https://doi.org/10.1016/bs.mie.2018.09.025