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Characterization of p-azidophenylalanine as a system L substrate and a photoaffinity probe

Journal Article · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:6989965
; ;
p-Azidophenylalanine (azphe) is a photosensitive tyrosine analog that produces a highly reactive nitrene upon UV irradiation. The authors have shown for the cultured hepatoma cell (HTC) that azphe is a specific substrate for System L. It shows competitive inhibition for the transport of the bicyclooctane amino acid analog BCO, leucine and other substrates for System L. It does not inhibit the Na/sup +/-dependent transport of MeAIB by System A, or the Na/sup +/-dependent uptake of alanine, serine and threonine by System ASC. In addition the Na/sup +/-independent uptake of arginine by System y/sup +/ was unaffected. Na/sup +/-independent kinetics indicate that the K/sub m/ of azphe (78 ..mu..M) is similar to that of other System L substrates. The K/sub i/'s of tryptophan, phenylalanine, and the norbornane amino acid analog, BCH, on azphe are also in the range of 75 to 120 /sup +/M. Upon UV irradiation with azphe, in both intact HTC cells and in HTC plasma membrane vesicles, the uptake of leucine becomes irreversibly inhibited. This irreversible inhibition is protected by the addition of 10 mM leucine but not 10 mM MeAIB. The authors are currently applying this procedure with tritiated azphe to identify the System L transport protein in plasma membranes from HTC cells.
Research Organization:
Univ. of Michigan Medical School, Ann Arbor
OSTI ID:
6989965
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

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Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACIDS
ANIMAL CELLS
AZIDO COMPOUNDS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL MEMBRANES
ELECTROMAGNETIC RADIATION
INHIBITION
LEUCINE
MEMBRANE TRANSPORT
MEMBRANES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHENYLALANINE
RADIATIONS
RADIOSENSITIVITY
SUBSTRATES
TUMOR CELLS
ULTRAVIOLET RADIATION