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Insertion and glycosylation of Pf3‐derived membrane proteins in microsomes
Author(s) -
Ridder Anja,
Thissen Laura,
Killian Antoinette,
de Kruijff Ben
Publication year - 2002
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/s0014-5793(02)02312-8
Subject(s) - glycosylation , insert (composites) , vesicle , biochemistry , chemistry , membrane protein , membrane , mutant , escherichia coli , microsome , amino acid , peptide sequence , biophysics , in vitro , biology , gene , mechanical engineering , engineering
To get insight into the insertion mechanism of small newly synthesized single‐spanning membrane proteins, Pf3 coat protein mutants were constructed with potential glycosylation sites in the N‐terminus. Some of these proteins, when synthesized in vitro in the presence of microsomes, became efficiently glycosylated, proving that they insert into the membrane and translocate their N‐terminus to the lumenal side. Such Pf3 constructs also insert efficiently into Escherichia coli vesicles and even in pure lipid vesicles, suggesting a common mechanism, which might be spontaneous. Glycosylation was sensitive to changes in the amino acid sequence of the N‐terminus, suggesting that it depends on the structure of the protein and/or its positioning with respect to the lipid–water interface.