Abstract

Comparison of subunit sequences and X-ray crystal structures of ribulose-1,5-bisphosphate carboxylase/oxygenase indicates that the loop between beta-strands A and B of the small subunit is one of the most variable regions of the holoenzyme. In prokaryotes and nongreen algae, the loop contains 10 residues. In land plants and green algae, the loop is comprised of similar to 22 and 28 residues, respectively. Previous studies indicated that the longer beta A-beta B loop was required for the assembly of cyanobacterial small subunits with plant large subunits in isolated chloroplasts. In the present study, chimeric small subunits were constructed by replacing the loop of the green alga Chlamydomonas reinhardtii with the sequences of Synechococcus or spinach. When these engineered genes were transformed into a Chlamydomonas mutant that lacks small-subunit genes, photosynthesis-competent colonies were recovered, indicating that loop size is not essential for holoenzyme assembly. Whereas the Synechococcus loop causes decreases in carboxylation V-max, K-m(O-2), and CO2/O-2 specificity, the spinach loop causes complementary decreases in carboxylation V-max, K-m(O-2), and K-m(CO2) without a change in specificity. X-ray crystal structures of the engineered proteins reveal remarkable similarity between the introduced beta A-beta B loops and the respective loops in the Synechococcus and spinach enzymes. The side chains of several large-subunit residues are altered in regions previously shown by directed mutagenesis to influence CO2/O-2 specificity. Differences in the catalytic properties of divergent Rubisco enzymes may arise from differences in the small-subunit beta A-beta B loop. This loop may be a worthwhile target for genetic engineering aimed at improving photosynthetic CO2 fixation.

Published in

Biochemistry
2005, Volume: 44, number: 29, pages: 9851-9861
Publisher: AMER CHEMICAL SOC

SLU Authors

• Karkehabadi, Saeid

  • Department of Molecular Biology, Swedish University of Agricultural Sciences
    

• Taylor, Tom

  • Department of Molecular Biology, Swedish University of Agricultural Sciences
    

• Andersson, Inger

  • Department of Molecular Biology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Structural Biology
Biochemistry and Molecular Biology
Forest Science


Publication identifier

DOI: https://doi.org/10.1021/bi050537v

Permanent link to this page (URI)

https://res.slu.se/id/publ/66083