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Sequence of a benzyladenine binding site peptide isolated from a wheat embyro cytokinin-binding protein

Conference · · Plant Physiol., Suppl.; (United States)
OSTI ID:5653449
; ; ; ; ;

A wheat embryo cytokinin-binding protein (CBF-1) was covalently modified with the radiolabeled photoaffinity ligand (/sup 14/C)-2-azido-N/sup 6/-benzyladenine (AzBA). A single labeled peptide was obtained after proteolytic digestion and isolation by reversed-phase and anion exchange HPLC. Sequencing by Edman degradation identified 11 of the 12 residues, but failed to identify the labeled amino acid. Amino acid analysis and sequencing by laser photodissociation-Fourier transform mass spectrometry conclusively identified the labeled residue as the histidine: Ala-Phe-Leu-Gln-Pro-Ser-His-His*-Asp-Ala-Asp-Glu. Comparison of this sequence with the known partial primary sequence of CBF-1 (determined by tandem quadrupole mass spectrometry) showed that there were two homologous sequences in CBF-1. One exactly matched the above AzBA labeled peptide and the other differed by a substitution of tyrosine for the second histidine. The possibility of binding and non-binding CBF-1 isomers will be discussed along with a proposed model for BA binding.

Research Organization:
Cell Research Institute, Dublin, CA
OSTI ID:
5653449
Report Number(s):
CONF-8707108-
Journal Information:
Plant Physiol., Suppl.; (United States), Journal Name: Plant Physiol., Suppl.; (United States) Vol. 83:4; ISSN PPYSA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CEREALS
CHROMATOGRAPHY
DIGESTION
ELECTROMAGNETIC RADIATION
GRASS
ISOTOPE APPLICATIONS
KINETICS
KININS
LABELLED COMPOUNDS
LASER RADIATION
LIGANDS
LIQUID COLUMN CHROMATOGRAPHY
MASS SPECTROSCOPY
MOLECULAR STRUCTURE
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDES
PLANT GROWTH REGULATORS
PLANTS
POLYPEPTIDES
PROTEINS
RADIATIONS
REACTION KINETICS
SEPARATION PROCESSES
SPECTROSCOPY
TRACER TECHNIQUES
WHEAT