Tic20 and Tic22 Are New Components of the Protein Import Apparatus at the Chloroplast Inner Envelope Membrane

doi:10.1083/jcb.143.4.991

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© The Rockefeller University Press, 0021-9525/1998//991 $5.00
The Journal of Cell Biology, Volume 143, Number 4, , 1998 991-1002

Regular Articles

Tic20 and Tic22 Are New Components of the Protein Import Apparatus at the Chloroplast Inner Envelope Membrane

Andrei Kouranov, Xuejun Chen, Bruno Fuks, and Danny J. Schnell

Department of Biological Sciences, Rutgers, The State University of New Jersey, Newark, New Jersey 07102

Two components of the chloroplast envelope, Tic20 and Tic22,were previously identified as candidates for components ofthe general protein import machinery by their ability to covalentlycross-link to nuclear-encoded preproteins trapped at an intermediatestage in import across the envelope (Kouranov, A., and D.J.Schnell. 1997. J. Cell Biol. 139:1677–1685). We havedetermined the primary structures of Tic20 and Tic22 and investigatedtheir localization and association within the chloroplast envelope.Tic20 is a 20-kD integral membrane component of the inner envelope membrane. In contrast, Tic22 is a 22-kD protein that is locatedin the intermembrane space between the outer and inner envelopemembranes and is peripherally associated with the outer faceof the inner membrane. Tic20, Tic22, and a third inner membraneimport component, Tic110, associate with import components of the outer envelope membrane. Preprotein import intermediatesquantitatively associate with this outer/inner membrane supercomplex,providing evidence that the complex corresponds to envelopecontact sites that mediate direct transport of preproteins fromthe cytoplasm to the stromal compartment. On the basis of theseresults, we propose that Tic20 and Tic22 are core componentsof the protein translocon of the inner envelope membrane ofchloroplasts.

Key Words: chloroplast • protein import • membrane translocation • protein targeting

Abbreviations used in this paper: EST, expressed sequence tag; protA, IgG-binding domain of staphylococcal protein A; pS, precursor of the small subunit of ribulose-1,5-bisphosphate carboxylase; Tic, translocon at the inner envelope membrane of chloroplasts; Toc, translocon at the outer envelope membrane of chloroplasts.

A. Kouranov and X. Chen contributed equally to this work.

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