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Identification of a putative membrane‐interacting domain of CTP: Phosphocholine cytidylyltransferase from rat liver
Author(s) -
Wieder Thomas,
Geilen Christoph C.,
Wieprecht Marcus,
Becker Andreas,
Orfanos Constantin E.
Publication year - 1994
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(94)00433-1
Subject(s) - phosphocholine , identification (biology) , chemistry , biochemistry , domain (mathematical analysis) , membrane , microbiology and biotechnology , biophysics , biology , phospholipid , phosphatidylcholine , mathematical analysis , botany , mathematics
A putative membrane‐interacting domain of CTP:phosphocholine cytidylyltransferase (CT) was identified using two peptide‐specific antibodies. One antibody (SA2) was raised against the N‐terminus of CT (amino acid residues 1–17) and the other antibody (SA209) against an α‐helical domain of the enzyme (amino acid residues 247–257). Both antibodies quantitatively immunoprecipitated CT from rat liver cytosol and showed specificity towards CT when octylglucoside extracts of rat liver cytosol were assessed by Western blot analysis. However, further experiments revealed that the antibodies had different characteristics. Whereas the antibody directed against the N‐terminus of CT (SA2) did not influence CT/membrane interaction, the new antibody (SA209) against the α‐helical domain of the enzyme interfered with this interaction. Our results provide experimental evidence that the α‐helical domain (amino acid residues 228–287) of CT may serve as a membrane‐interacting domain.