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J Biol Chem, Vol. 274, Issue 8, 4715-4721, February 19, 1999
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From the The biogenesis of several bacterial polytopic
membrane proteins has been shown to require signal recognition particle
(SRP) and protein transport machinery, and one such protein, the major light-harvesting chlorophyll-binding protein (LHCP) exhibits these requirements in chloroplasts. In this report we have used in
vitro insertion assays to analyze four additional members of the
chlorophyll-a/b-binding protein family. We show
that two members, Lhca1 and Lhcb5, display an absolute requirement for
stroma, nucleoside triphosphates, and protein transport apparatus,
indicating an "assisted" pathway that probably resembles that of
LHCP. Two other members, however, namely an early light-inducible
protein 2 (Elip2) and photosystem II subunit S (PsbS), can insert
efficiently in the complete absence of SRP, SecA activity, nucleoside
triphosphates, or a functional Sec system. The data suggest a possibly
spontaneous insertion mechanism that, to date, has been characterized
only for simple single-span proteins. Of the membrane proteins whose
insertion into thylakoids has been analyzed, five have now been shown
to insert by a SRP/Sec-independent mechanism, suggesting that this is a
mainstream form of targeting pathway. We also show that PsbS and Elip2
molecules are capable of following either "unassisted" or assisted
pathways, and we discuss the implications for the mechanism and role of
SRP in chloroplasts.
Department of Biological Sciences,
University of Warwick, Coventry CV4 7AL, United Kingdom, the
§ Department of Plant Physiology, University of Umeå,
901 87 Umeå, Sweden, and the ¶ Department of Plant Biology,
Carnegie Institute of Washington, Stanford, California 94305
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