Site on the human erythrocyte glucose transporter phosphorylated by protein kinase C resides on the protein's hydrophilic domain

Journal Article · Thu May 01 04:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:7025314

Deziel, M R; McReynolds, J H; Lippes, H A; Jung, C Y

A recently published model of the human erythrocyte hexose transporter deduced from the protein's primary structure proposes that the transporter is organized into two membrane domains comprising 77% of the protein's mass and three hydrophilic domains, a short segment that includes the polypeptide's N-terminus and two larger segments, one lying between the membrane domains and the other at the protein's C-terminus. Limited tryptic digestion of the transporter produces two membrane-bound fragments corresponding to the proposed membrane domains and releases a number of soluble peptides. Fast Atom Bombardment Mass Spectroscopic analysis of the released peptides and comparison of the peptide's masses with the transporter's amino acid sequence revealed that tryptic peptides corresponding to at least 63% of the hydrophilic domains' mass were recovered. The site of phosphorylation by protein kinase C, tagged using (/sup 32/P)-ATP, was also released from the transporter under these conditions, (in contrast to sites located within the protein's membrane domains), indicating that this site is located within one of the hydrophilic domains. Tryptic digestion at elevated ionic strength or cleavage with S. Aureus V8 protease results in the recovery of the /sup 32/P label on the carbohydrate-bearing membrane domain that is located near the protein's N-terminus, thus eliminating the C-terminal hydrophilic segment as a possible site of phosphorylation.

• Research Organization: State Univ. of New York, Buffalo
• OSTI ID: 7025314
• Report Number(s): CONF-8606151-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMALS
ATP
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
ENZYMES
ERYTHROCYTES
GLUCOSE
HEXOSES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAN
MATERIALS
MEMBRANE TRANSPORT
MEMBRANES
MOLECULAR STRUCTURE
MONOSACCHARIDES
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PRIMATES
PROTEINS
RADIOISOTOPES
SACCHARIDES
TRACER TECHNIQUES
TRANSFERASES
VERTEBRATES