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S ‐( p ‐Azidophenacyl)thiocarnitine specifically binds to the γ‐butyrobetaine‐binding protein of Agrobacterium sp. and can be used as a photoaffinity label
Author(s) -
Nobile S.,
Baccino D.,
Deshusses J.
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80454-x
Subject(s) - reagent , photoaffinity labeling , periplasmic space , chemistry , stereochemistry , agrobacterium , biochemistry , binding protein , binding site , escherichia coli , transformation (genetics) , organic chemistry , gene
The photoaffinity reagent S ‐( p ‐azidophenacyl)thiocarnitine (PAP‐TC) has been synthesized according to Mauro et al. [(1986) Biochem. J. 237, 533–540]. This compound, originally designed for a structure‐function study of carnitine acetyl‐transferase, was used to analyze the Agrobacterium sp. γ‐butyrobetaine transport system. PAP‐TC appears to be a reagent specific to the transport system since it showed a competitive inhibition ( K i = 70 μM) of γ‐butyrobetaine transport. UV irradiation of periplasmic proteins in the presence of [ 14 C]PAP‐TC resulted in the irreversible labeling of the γ‐butyrobetaine‐binding protein. The addition of 1 mM γ‐butyrobetaine in the mixture significantly decreased the incorporation of the reagent, showing that this compound reacts specifically with the binding protein.