Distant downstream sequence determinants can control N-tail translocation during protein insertion into the endoplasmic reticulum membrane
2000 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 275, no 9, 6207-6213 p.Article in journal (Refereed) Published
We have studied the membrane insertion of ProW, an Escherichia coli inner membrane protein with seven transmembrane segments and a large periplasmic N-terminal tail, into endoplasmic reticulum (ER)-derived dog pancreas microsomes. Strikingly, significant levels of N-tail translocation is seen only when a minimum of four of the transmembrane segments are present; for constructs with fewer transmembrane segments, the N-tail remains mostly nontranslocated and the majority of the molecules adopt an "inverted" topology where normally nontranslocated parts are translocated and vice versa, N-tail translocation can also be promoted by shortening of the N-tail and by the addition of positively charged residues immediately downstream of the first transmembrane segment. We conclude that as many as four consecutive transmembrane segments may be collectively involved in determining membrane protein topology in the ER and that the effects of downstream sequence determinants may vary depending on the size and charge of the N-tail, We also provide evidence to suggest that the ProW N-tail is translocated across the ER membrane in a C-to-N-terminal direction.
Place, publisher, year, edition, pages
2000. Vol. 275, no 9, 6207-6213 p.
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:su:diva-21426DOI: 10.1074/jbc.275.9.6207ISI: 000085654400026PubMedID: 10692414OAI: oai:DiVA.org:su-21426DiVA: diva2:187953