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Structural analysis of the messenger RNA cap-binding protein: presence of phosphate, sulfhydryl, and disulfide groups

Conference · · J. Biol. Chem.; (United States)
OSTI ID:5150816
The messenger-RNA cap-binding protein (CBP) was isolated from human erythrocyte, rabbit erythrocyte, and rabbit reticulocyte lysate by affinity chromatography on 7-methylguanosine 5'-triphosphate-Sepharose. The specific activity of binding to capped oligonucleotides was similar for the human erythrocyte and rabbit reticulocyte CBPs. Isoelectric focusing of human and rabbit preparations revealed that each was composed of up to five species. The pI values of human and rabbit CBPs ranged from 5.7 to 6.5. The predominant form in erythrocytes had a pI of 6.3 while in reticulocytes, two major species, having pI values of 5.9 and 6.3, were present. Labeling of rabbit reticulocytes with (/sup 32/P)orthophosphate revealed that the pI 5.9 but not the pI 6.3 form contained phosphate. All of the phosphate was found in phosphoserine residues. The amino acid compositions of human erythrocyte and rabbit reticulocyte CBPs were quite similar. Both proteins had 7 tryptophanyl and 6 cysteinyl residues. Labeling with (1-/sup 14/C)iodoacetic acid under native and denaturing conditions provided evidence that 2 of the cysteinyl residues are present in the reduced form and 4 in disulfide bridges.
Research Organization:
Univ. of Kentucky, Lexington
OSTI ID:
5150816
Conference Information:
Journal Name: J. Biol. Chem.; (United States) Journal Volume: 261:1
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
AROMATICS
AZAARENES
AZOLES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CHROMATOGRAPHY
CYSTEINE
DATA
DAYS LIVING RADIOISOTOPES
ERYTHROCYTES
EXPERIMENTAL DATA
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
INDOLES
INFORMATION
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
MAMMALS
MATERIALS
MESSENGER-RNA
NUCLEI
NUCLEIC ACIDS
NUMERICAL DATA
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PROTEIN STRUCTURE
PROTEINS
PYRROLES
RABBITS
RADIOISOTOPES
REACTION KINETICS
RETICULOCYTES
RNA
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
THIOLS
TRACER TECHNIQUES
TRYPTOPHAN
VERTEBRATES