Extended Data Fig. 3: PAGE analyses.
From: Novel bacterial clade reveals origin of form I Rubisco
a, Non-denaturing PAGE gel with a molecular weight marker (M, lane 1), and purified proteins of all three candidate form I' Rubisco (P. breve, 241187, and 170907) with (+) or without (-) prior activation and incubation with 10-fold molar excess of 2CABP. 241187 and 170907 denotes scaffolds B_1_S1_170907_scaffold_241187_5_Tax=RBG_16_Chloroflexi_63_12 and S_p2_S4_170907_scaffold_85440 Rubisco, respectively. b, SDS–PAGE analysis of crude cell lysate from 1) overexpression of untagged P. breve Rubisco with co-expression of GroEL/ES from pBAD33EL/ES, 2) overexpression of His14-bdSUMO-tagged P. breve Rubisco with co-expression of GroEL/ES from pBAD33EL/ES, and 3) overexpression of His14-bdSUMO-tagged P. breve Rubisco without overexpression of GroEL/ES (background GroEL/ES expression from E. coli). Without GroEL/ES overexpression, untagged RbcL comprises 8 ± 1.0 (n = 3) of the total soluble protein, which improves to 14 ± 0.5 (n = 3) when GroEL/ES overexpression in induced (see Methods). When the His14-bdSUMO tag is included on the N-terminal end of RbcL, soluble expression is 7 ± 0.8 (n = 3) and 14 ± 0.8 (n = 3) of the total soluble protein, without and with GroEL/ES overexpression, respectively. Reported values collected from n separate experiments (separately grown E. coli cultures) reflect the mean ± standard deviation.
