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Chlorophyll a/b binding (CAB) polypeptides of CP29, the internal chlorophyll a/b complex of PSII: characterization of the tomato gene encoding the 26 kDa (type 1) polypeptide, and evidence for a second CP29 polypeptide

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Summary

CP29, the core chlorophyll a/b (CAB) antenna complex of Photosystem II (PSII), has two nuclearencoded polypeptides of approximately 26 and 28 kDa in tomato (Lycopersicon esculentum). Cab9, the gene for the Type 1 (26 kDa) CP29 polypeptide was cloned by immunoscreening a tomato leaf cDNA library. Its identity was confirmed by sequencing tryptic peptides from the mature protein. Cab9 is a single-copy gene with five introns, the highest number found in a CAB protein. In vitro transcription-translation gave a 31 kDa precursor which was cleaved to about 26 kDa after import into isolated tomato chloroplasts. The Cab9 polypeptide has the two highly conserved regions common to all CAB polypeptides, which define the members of this extended gene family. Outside of the conserved regions, it is only slightly more closely related to other PSII CABs than to PSI CABs. Sequence analysis of tryptic peptides from the Type II (28 kDa) CP29 polypeptide showed that it is also a member of the CAB family and is very similar or identical to the CP29 polypeptide previously isolated from spinach. All members of the CAB family have absolutely conserved His, Gln and Asn residues which could ligate the Mg atoms of the chlorophylls, and a number of conserved Asp, Glu, Lys and Arg residues which could form H-bonds to the polar groups on the porphyrin rings. The two conserved regions comprise the first and third predicted trans-membrane helices and the stroma-exposed segments preceding them.

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Abbreviations

Chl:

chlorophyll

CP29:

core Chl a/b antenna of Photosystem II

LHCII:

major Chl a/b antenna of Photosystem II

LHCl:

Chl a/b antenna of Photosystem 1

PS:

Photosystem

CPII* :

oligomeric form of LHCII isolated by SDS-PAGE

SDSPAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

HPLC:

high-performance liquid chromatography

TMH:

transmembrane (membrane-spanning) helix

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Authors and Affiliations

  1. Biology Department, University of Michigan, 48109, Ann Arbor, MI, USA

    Eran Pichersky & James Reid

  2. Botany Department, University of British Columbia, V6T 2B1, Vancouver, B.C., Canada

    Rajagopal Subramaniam & Beverley R. Green

  3. Botany Department, N. Carolina State University, Raleigh, N.C., USA

    Michael J. Whites

  4. Biomedical Research Centre and Biochemistry Dept., University of British Columbia, V6T 1W5, Vancouver, B.C., Canada

    Ruedi Aebersold

Authors
  1. Eran Pichersky
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  2. Rajagopal Subramaniam
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  3. Michael J. Whites
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  4. James Reid
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  5. Ruedi Aebersold
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  6. Beverley R. Green
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Communicated by R.G. Herrmann

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Pichersky, E., Subramaniam, R., Whites, M.J. et al. Chlorophyll a/b binding (CAB) polypeptides of CP29, the internal chlorophyll a/b complex of PSII: characterization of the tomato gene encoding the 26 kDa (type 1) polypeptide, and evidence for a second CP29 polypeptide. Molec. Gen. Genet. 227, 277–284 (1991). https://doi.org/10.1007/BF00259681

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