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. 2016 Dec 1;113(51):14716–14721. doi: 10.1073/pnas.1616294113

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Fig. 1. — Purification and characterization of LCIB homologs. (A) SDS/PAGE analysis. (B) Gel filtration analysis of recombinant CrLCIB and CrLCIC complexes. LCIB-LCIC was prepared by recombinantly coexpressing CrLCIB and CrLCIC. LCIB+LCIC was prepared by mixing and incubating separately expressed CrLCIB and CrLCIC in vitro. In each experiment, a total of 25 μg per sample was loaded, and the corresponding fractions were separated by SDS/PAGE and are shown below the chromatograph. (C) Gel filtration analysis of diatom and prokaryotic LCIB homologs, with 50 μg per isoform loaded in each experiment. (D) MIMS CA activity assay for LCIB homologs reported as the first-order rate constant for the hydration of CO₂ (k_CA), μmol mg protein⁻¹ s⁻¹ μM CO₂⁻¹ at 25 °C. LCIB-LCIC (native), CrLCIB-LCIC complex purified from C. reinhardtii cells. (E) Wilbur–Anderson CA activity assay of the LCIB proteins. Error bars indicate the mean and SD of at least three independent repeats.