CC-4942 rbcL-C256F mt+ (new)
$30.00
From Robert J. Spreitzer, University of Nebraska, November 2014
Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a C256F substitution (TGT-TTC) was created in the Rubisco large subunit. The same substitution was created previously in a different host strain (see CC-4839) (Du et al. 2003). Because Cys-256 is methylated in Chlamydomonas Rubisco (Taylor et al. 2001), but replaced by Phe in plant Rubisco (Du et al. 2003), the mutant was created to investigate the role of the modified residue in Rubisco structure or function. The C256F substitution causes a small decrease in carboxylation catalytic efficiency (Du et al. 2003), but does not affect the photosynthetic growth of the mutant cells (Spreitzer et al., unpublished). C256F has been studied in combination with other phylogenetic substitutions (Du et al. 2003; Spreitzer et al. 2005). See also rbcL-C256F/C369V mt+. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.
Du YC, Peddi SR, Spreitzer RJ (2003) Assessment of structural and functional divergence far from the large subunit active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 278:49401-49405
Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244
Spreitzer RJ, Peddi SR, Satagopan S (2005) Phylogenetic engineering at an interface between large and small subunits imparts land-plant kinetic properties to algal Rubisco. Proc Natl Acad Sci USA 102:17225-17230
Taylor TC, Backlund A, Bjorhall K, Spreitzer RJ, Andersson I (2001) First crystal structure of Rubisco from a green alga, Chlamydomonas reinhardtii. J Biol Chem 276:48159-48164