Sec-independent insertion of thylakoid membrane proteins: Analysis of insertion forces and identification of a loop intermediate involving the signal peptide

Simon J. Thompson, Soo Jung Kim, Colin Robinson

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A group of membrane proteins are synthesized with cleavable signal sequences but inserted into the thylakoid membrane by an unusual Sec/SRP- independent mechanism. In this report we describe a key intermediate in the insertion of one such protein, photosystem II subunit W (PSII-W). A single mutation in the terminal cleavage site partially blocks processing and leads to the formation of an intermediate-size protein in the thylakoid membrane during chloroplast import assays. This protein is in the form of a loop structure: the N and C termini are exposed on the stromal face, whereas the cleavage site has been translocated into the lumen. In this respect the insertion of this protein resembles that of M13 procoat, which also adopts a loop structure during insertion, and we present preliminary evidence that a similar mechanism is used by another thylakoid protein, PSII-X. However, whereas the negatively charged region of procoat is translocated by an apparently electrophoretic mechanism using the ΔμH +, the corresponding region of PSII-W is equally acidic but insertion is ΔμH + independent. We furthermore show that neutralization of this region has no apparent effect on the insertion process. We propose that a central element in this insertion mechanism is a loop structure whose formation is driven by hydrophobic interactions.

Original languageEnglish (US)
Pages (from-to)18979-18983
Number of pages5
JournalJournal of Biological Chemistry
Volume273
Issue number30
DOIs
StatePublished - Jul 24 1998
Externally publishedYes

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Thylakoid Membrane Proteins
Protein Sorting Signals
Thylakoids
Photosystem II Protein Complex
Proteins
Membranes
Chloroplasts
Hydrophobic and Hydrophilic Interactions
Membrane Proteins
Assays
Mutation
Processing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sec-independent insertion of thylakoid membrane proteins : Analysis of insertion forces and identification of a loop intermediate involving the signal peptide. / Thompson, Simon J.; Kim, Soo Jung; Robinson, Colin.

In: Journal of Biological Chemistry, Vol. 273, No. 30, 24.07.1998, p. 18979-18983.

Research output: Contribution to journalArticle

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