Radioiodination of cell-surface glycoproteins by carbohydrate modification
Mild oxidation of cell-surface sialic acid residues followed by reduction with sodium /sup 3/H-borohydride is a common method of radiolabeling glycoproteins. In many cases it is desirable to incorporate into glycoproteins a label of higher specific activity such as /sup 125/I. Incorporation of modified amino compounds into oxidized, isolated glycoproteins by reductive amination has been demonstrated by several investigators. They have determined the conditions for the application of this approach to radioiodination of intact cells. Cells are oxidized by exposure to 1 mM sodium periodate. Tyrosine or a tyrosine derivative, radiolabeled to high specific activity with Iodogen and carrier-free Na/sup 125/I, is added, followed by 1 mM sodium cyanoborohydride. Labeled cell-surface proteins are analyzed by SDS-gel electrophoresis of cell lysates. The addition of excess carrier glycoprotein, such as fetuin, is necessary to prevent degradation of the labeled product in the cell lysate. The incorporation of radiolabel can approach that of direct iodination of cell-surface tyrosyl residues, about 100 dpm/cell. The labeling procedure has been applied to the analysis of murine lymphocyte glycoproteins.
- Research Organization:
- Univ. of Texas Health Science Center, San Antonio
- OSTI ID:
- 5021814
- Report Number(s):
- CONF-8606151-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLUCOPROTEINS
LABELLING
ELECTROPHORESIS
IODINE 125
RESIDUES
SIALIC ACID
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TYROSINE
AMINO ACIDS
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
HYDROXY ACIDS
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
MONOSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
RADIOISOTOPES
SACCHARIDES
550201* - Biochemistry- Tracer Techniques