Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor

Taysir K. Bader; Todd M. Rappe; Gianlugi Veglia; Mark D. Distefano

doi:10.1016/bs.mie.2018.09.025

Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor

Taysir K. Bader, Todd M. Rappe, Gianlugi Veglia, Mark D. Distefano

Research output: Chapter in Book/Report/Conference proceeding › Chapter

2 Scopus citations

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.

Original language	English (US)
Title of host publication	Biological NMR Part A
Editors	A. Joshua Wand
Publisher	Academic Press Inc.
Pages	207-238
Number of pages	32
Volume	614
ISBN (Print)	9780128138601
DOIs	https://doi.org/10.1016/bs.mie.2018.09.025
State	Published - Jan 1 2019

Publication series

Name	Methods in enzymology
Publisher	Academic Press Inc.
ISSN (Print)	0076-6879

Bibliographical note

Funding Information:
Mass spectrometry analysis was performed at The University of Minnesota Department of Chemistry Mass Spectrometry Laboratory (MSL), supported by the Office of the Vice President of Research, College of Science and Engineering, and the Department of Chemistry at the University of Minnesota, as well as The National Science Foundation (NSF, Award CHE-1336940). The content of this chapter is the sole responsibility of the authors and does not represent endorsement by the MSL or NSF. This work was supported in part by the National Institutes of Health (RF1AG056976, GM084152, and GM106082) and by the National Science Foundation (CHE-1308655). NMR analysis was performed at Minnesota NMR center. Funding for NMR instrumentation was provided by the Office of the Vice President for Research, the Medical School, the College of Biological Science, NIH, NSF, and the Minnesota Medical Foundation.

Publisher Copyright:
© 2019 Elsevier Inc.

Keywords

C-terminal ester
Farnesylation
Lipidation
NMR
Peptide
Pheromone
Prenylation
a-Factor
Mating Factor/chemical synthesis
Humans
Fluorenes/chemistry
Trityl Compounds/chemistry
Nuclear Magnetic Resonance, Biomolecular/methods
Protein Prenylation
Solid-Phase Synthesis Techniques/methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
Peptides/chemical synthesis
Chromatography, Affinity/methods
Saccharomyces cerevisiae/chemistry

PubMed: MeSH publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Journal Article
Research Support, N.I.H., Extramural

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/bs.mie.2018.09.025

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title = "Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor",

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 = "C-terminal ester, Farnesylation, Lipidation, NMR, Peptide, Pheromone, Prenylation, a-Factor, Mating Factor/chemical synthesis, Humans, Fluorenes/chemistry, Trityl Compounds/chemistry, Nuclear Magnetic Resonance, Biomolecular/methods, Protein Prenylation, Solid-Phase Synthesis Techniques/methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods, Peptides/chemical synthesis, Chromatography, Affinity/methods, Saccharomyces cerevisiae/chemistry",

author = "Bader, {Taysir K.} and Rappe, {Todd M.} and Gianlugi Veglia and Distefano, {Mark D.}",

note = "Funding Information: Mass spectrometry analysis was performed at The University of Minnesota Department of Chemistry Mass Spectrometry Laboratory (MSL), supported by the Office of the Vice President of Research, College of Science and Engineering, and the Department of Chemistry at the University of Minnesota, as well as The National Science Foundation (NSF, Award CHE-1336940). The content of this chapter is the sole responsibility of the authors and does not represent endorsement by the MSL or NSF. This work was supported in part by the National Institutes of Health (RF1AG056976, GM084152, and GM106082) and by the National Science Foundation (CHE-1308655). NMR analysis was performed at Minnesota NMR center. Funding for NMR instrumentation was provided by the Office of the Vice President for Research, the Medical School, the College of Biological Science, NIH, NSF, and the Minnesota Medical Foundation. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",

year = "2019",

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T1 - Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor

AU - Bader, Taysir K.

AU - Rappe, Todd M.

AU - Veglia, Gianlugi

AU - Distefano, Mark D.

N1 - Funding Information: Mass spectrometry analysis was performed at The University of Minnesota Department of Chemistry Mass Spectrometry Laboratory (MSL), supported by the Office of the Vice President of Research, College of Science and Engineering, and the Department of Chemistry at the University of Minnesota, as well as The National Science Foundation (NSF, Award CHE-1336940). The content of this chapter is the sole responsibility of the authors and does not represent endorsement by the MSL or NSF. This work was supported in part by the National Institutes of Health (RF1AG056976, GM084152, and GM106082) and by the National Science Foundation (CHE-1308655). NMR analysis was performed at Minnesota NMR center. Funding for NMR instrumentation was provided by the Office of the Vice President for Research, the Medical School, the College of Biological Science, NIH, NSF, and the Minnesota Medical Foundation. Publisher Copyright: © 2019 Elsevier Inc.

PY - 2019/1/1

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N2 - 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.

AB - 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.

KW - C-terminal ester

KW - Farnesylation

KW - Lipidation

KW - NMR

KW - Peptide

KW - Pheromone

KW - Prenylation

KW - a-Factor

KW - Mating Factor/chemical synthesis

KW - Humans

KW - Fluorenes/chemistry

KW - Trityl Compounds/chemistry

KW - Nuclear Magnetic Resonance, Biomolecular/methods

KW - Protein Prenylation

KW - Solid-Phase Synthesis Techniques/methods

KW - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods

KW - Peptides/chemical synthesis

KW - Chromatography, Affinity/methods

KW - Saccharomyces cerevisiae/chemistry

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U2 - 10.1016/bs.mie.2018.09.025

DO - 10.1016/bs.mie.2018.09.025

M3 - Chapter

C2 - 30611425

AN - SCOPUS:85058776513

SN - 9780128138601

VL - 614

T3 - Methods in enzymology

SP - 207

EP - 238

BT - Biological NMR Part A

A2 - Wand, A. Joshua

PB - Academic Press Inc.

ER -

Synthesis and NMR Characterization of the Prenylated Peptide, a-Factor

Abstract

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Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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