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Title: Characterization of a benzyladenine binding-site peptide isolated from a wheat cytokinin-binding protein: Sequence analysis and identification of a single affinity-labeled histidine residue by mass spectrometry

Abstract

A wheat embryo cytokinin-binding protein was covalently modified with the radiolabeled photoaffinity ligand 2-azido-N{sup 6}-({sup 14}C)benzyladenine. A single labeled peptide was obtained after proteolytic digestion and isolation by reversed-phase and anion-exchange HPLC. Sequencing by classical Edman degradation identified 11 of the 12 residues but failed to identify the labeled amino acid. Analysis by laser photodissociation Fourier-transform mass spectrometry of 10 pmol of the peptide independently confirmed the Edman data and also demonstrated that the histidine residue nearest the C terminus (underlined) was modified by the reagent in the sequence Ala-Phe-Leu-Gln-Pro-Ser-His-His{und His}-Asp-Ala-Asp-Glu.

Authors:
; ; ; ; ; ;  [1]
  1. ARCO Plant Cell Research Institute, Dublin, CA (USA)
Publication Date:
OSTI Identifier:
5517444
Resource Type:
Journal Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America; (USA)
Additional Journal Information:
Journal Volume: 85:16; Journal ID: ISSN 0027-8424
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ADENINES; CROSS-LINKING; KININS; PROTEINS; MOLECULAR STRUCTURE; AMINO ACID SEQUENCE; CARBON 14 COMPOUNDS; HISTIDINE; ION EXCHANGE CHROMATOGRAPHY; LIGANDS; MASS SPECTROSCOPY; WHEAT; AMINES; AMINO ACIDS; ANTIMETABOLITES; AROMATICS; AZAARENES; AZOLES; CARBOXYLIC ACIDS; CEREALS; CHEMICAL REACTIONS; CHROMATOGRAPHY; DRUGS; GRAMINEAE; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; IMIDAZOLES; LABELLED COMPOUNDS; LILIOPSIDA; MAGNOLIOPHYTA; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PEPTIDES; PLANTS; POLYMERIZATION; POLYPEPTIDES; PURINES; SEPARATION PROCESSES; SPECTROSCOPY; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Brinegar, A C, Cooper, G, Stevens, A, Hauer, C R, Shabanowitz, J, Hunt, D F, and Fox, J E. Characterization of a benzyladenine binding-site peptide isolated from a wheat cytokinin-binding protein: Sequence analysis and identification of a single affinity-labeled histidine residue by mass spectrometry. United States: N. p., 1988. Web. doi:10.1073/pnas.85.16.5927.
Brinegar, A C, Cooper, G, Stevens, A, Hauer, C R, Shabanowitz, J, Hunt, D F, & Fox, J E. Characterization of a benzyladenine binding-site peptide isolated from a wheat cytokinin-binding protein: Sequence analysis and identification of a single affinity-labeled histidine residue by mass spectrometry. United States. doi:10.1073/pnas.85.16.5927.
Brinegar, A C, Cooper, G, Stevens, A, Hauer, C R, Shabanowitz, J, Hunt, D F, and Fox, J E. Mon . "Characterization of a benzyladenine binding-site peptide isolated from a wheat cytokinin-binding protein: Sequence analysis and identification of a single affinity-labeled histidine residue by mass spectrometry". United States. doi:10.1073/pnas.85.16.5927.
@article{osti_5517444,
title = {Characterization of a benzyladenine binding-site peptide isolated from a wheat cytokinin-binding protein: Sequence analysis and identification of a single affinity-labeled histidine residue by mass spectrometry},
author = {Brinegar, A C and Cooper, G and Stevens, A and Hauer, C R and Shabanowitz, J and Hunt, D F and Fox, J E},
abstractNote = {A wheat embryo cytokinin-binding protein was covalently modified with the radiolabeled photoaffinity ligand 2-azido-N{sup 6}-({sup 14}C)benzyladenine. A single labeled peptide was obtained after proteolytic digestion and isolation by reversed-phase and anion-exchange HPLC. Sequencing by classical Edman degradation identified 11 of the 12 residues but failed to identify the labeled amino acid. Analysis by laser photodissociation Fourier-transform mass spectrometry of 10 pmol of the peptide independently confirmed the Edman data and also demonstrated that the histidine residue nearest the C terminus (underlined) was modified by the reagent in the sequence Ala-Phe-Leu-Gln-Pro-Ser-His-His{und His}-Asp-Ala-Asp-Glu.},
doi = {10.1073/pnas.85.16.5927},
journal = {Proceedings of the National Academy of Sciences of the United States of America; (USA)},
issn = {0027-8424},
number = ,
volume = 85:16,
place = {United States},
year = {1988},
month = {8}
}