Matrix-assisted laser desorption/ionization of noncovalently bound compounds

A. S. Woods, J. C. Buchsbaum, T. A. Worrall, J. M. Berg, Robert J. Cotter

Research output: Contribution to journalArticle

Abstract

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to obtain spectra of peptide-metal ion complexes formed by a zinc finger peptide of the transcription factor IIIA (Cys2-His2) type and zinc and cobalt ions, as well as peptide-enzyme complexes. Peptides and proteins analyzed by mass spectrometry are generally dissolved in 0.1% aqueous TFA (pH <2.0). At this pH, peptides and proteins denature. We therefore reasoned that MALDI mass spectrometry might be able to detect noncovalently bound compounds if conditions were used to prepare the samples that allowed macromolecular assemblies to retain tertiary structure. Samples were dissolved in 1 M ammonium bicarbonate, and a saturated matrix solution was prepared using ethanol-ammonium bicarbonate (1:1) solution or ethanol-ammonium citrate (1:1) solution. All preparations of zinc finger peptides were done in a glovebox under nitrogen to prevent oxidation of the metal binding cysteine residues. Using this approach, we have been able to demonstrate that MALDI mass spectrometry can be used to study both noncovalent metal binding complexes and noncovalent peptide-enzyme complexes.

Original languageEnglish (US)
Pages (from-to)4462-4465
Number of pages4
JournalAnalytical Chemistry
Volume67
Issue number24
StatePublished - Dec 15 1995

Fingerprint

Ionization
Desorption
Peptides
Lasers
Mass spectrometry
Zinc
Transcription Factor TFIIIA
Ethanol
Metals
Enzymes
Cobalt
Metal ions
Cysteine
Proteins
Nitrogen
Ions
Oxidation

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Woods, A. S., Buchsbaum, J. C., Worrall, T. A., Berg, J. M., & Cotter, R. J. (1995). Matrix-assisted laser desorption/ionization of noncovalently bound compounds. Analytical Chemistry, 67(24), 4462-4465.

Matrix-assisted laser desorption/ionization of noncovalently bound compounds. / Woods, A. S.; Buchsbaum, J. C.; Worrall, T. A.; Berg, J. M.; Cotter, Robert J.

In: Analytical Chemistry, Vol. 67, No. 24, 15.12.1995, p. 4462-4465.

Research output: Contribution to journalArticle

Woods, AS, Buchsbaum, JC, Worrall, TA, Berg, JM & Cotter, RJ 1995, 'Matrix-assisted laser desorption/ionization of noncovalently bound compounds', Analytical Chemistry, vol. 67, no. 24, pp. 4462-4465.
Woods AS, Buchsbaum JC, Worrall TA, Berg JM, Cotter RJ. Matrix-assisted laser desorption/ionization of noncovalently bound compounds. Analytical Chemistry. 1995 Dec 15;67(24):4462-4465.
Woods, A. S. ; Buchsbaum, J. C. ; Worrall, T. A. ; Berg, J. M. ; Cotter, Robert J. / Matrix-assisted laser desorption/ionization of noncovalently bound compounds. In: Analytical Chemistry. 1995 ; Vol. 67, No. 24. pp. 4462-4465.
@article{307e0193bd84447ebaaa20b26993c3eb,
title = "Matrix-assisted laser desorption/ionization of noncovalently bound compounds",
abstract = "Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to obtain spectra of peptide-metal ion complexes formed by a zinc finger peptide of the transcription factor IIIA (Cys2-His2) type and zinc and cobalt ions, as well as peptide-enzyme complexes. Peptides and proteins analyzed by mass spectrometry are generally dissolved in 0.1{\%} aqueous TFA (pH <2.0). At this pH, peptides and proteins denature. We therefore reasoned that MALDI mass spectrometry might be able to detect noncovalently bound compounds if conditions were used to prepare the samples that allowed macromolecular assemblies to retain tertiary structure. Samples were dissolved in 1 M ammonium bicarbonate, and a saturated matrix solution was prepared using ethanol-ammonium bicarbonate (1:1) solution or ethanol-ammonium citrate (1:1) solution. All preparations of zinc finger peptides were done in a glovebox under nitrogen to prevent oxidation of the metal binding cysteine residues. Using this approach, we have been able to demonstrate that MALDI mass spectrometry can be used to study both noncovalent metal binding complexes and noncovalent peptide-enzyme complexes.",
author = "Woods, {A. S.} and Buchsbaum, {J. C.} and Worrall, {T. A.} and Berg, {J. M.} and Cotter, {Robert J.}",
year = "1995",
month = "12",
day = "15",
language = "English (US)",
volume = "67",
pages = "4462--4465",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "24",

}

TY - JOUR

T1 - Matrix-assisted laser desorption/ionization of noncovalently bound compounds

AU - Woods, A. S.

AU - Buchsbaum, J. C.

AU - Worrall, T. A.

AU - Berg, J. M.

AU - Cotter, Robert J.

PY - 1995/12/15

Y1 - 1995/12/15

N2 - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to obtain spectra of peptide-metal ion complexes formed by a zinc finger peptide of the transcription factor IIIA (Cys2-His2) type and zinc and cobalt ions, as well as peptide-enzyme complexes. Peptides and proteins analyzed by mass spectrometry are generally dissolved in 0.1% aqueous TFA (pH <2.0). At this pH, peptides and proteins denature. We therefore reasoned that MALDI mass spectrometry might be able to detect noncovalently bound compounds if conditions were used to prepare the samples that allowed macromolecular assemblies to retain tertiary structure. Samples were dissolved in 1 M ammonium bicarbonate, and a saturated matrix solution was prepared using ethanol-ammonium bicarbonate (1:1) solution or ethanol-ammonium citrate (1:1) solution. All preparations of zinc finger peptides were done in a glovebox under nitrogen to prevent oxidation of the metal binding cysteine residues. Using this approach, we have been able to demonstrate that MALDI mass spectrometry can be used to study both noncovalent metal binding complexes and noncovalent peptide-enzyme complexes.

AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to obtain spectra of peptide-metal ion complexes formed by a zinc finger peptide of the transcription factor IIIA (Cys2-His2) type and zinc and cobalt ions, as well as peptide-enzyme complexes. Peptides and proteins analyzed by mass spectrometry are generally dissolved in 0.1% aqueous TFA (pH <2.0). At this pH, peptides and proteins denature. We therefore reasoned that MALDI mass spectrometry might be able to detect noncovalently bound compounds if conditions were used to prepare the samples that allowed macromolecular assemblies to retain tertiary structure. Samples were dissolved in 1 M ammonium bicarbonate, and a saturated matrix solution was prepared using ethanol-ammonium bicarbonate (1:1) solution or ethanol-ammonium citrate (1:1) solution. All preparations of zinc finger peptides were done in a glovebox under nitrogen to prevent oxidation of the metal binding cysteine residues. Using this approach, we have been able to demonstrate that MALDI mass spectrometry can be used to study both noncovalent metal binding complexes and noncovalent peptide-enzyme complexes.

UR - http://www.scopus.com/inward/record.url?scp=0029569085&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029569085&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:0029569085

VL - 67

SP - 4462

EP - 4465

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 24

ER -