Peptide mapping of the ribulose bisphosphate carboxylase large subunit from the genus Oenothera

Abstract

A peptide mapping procedure for the peptides derived by chymotryptic digestion of the large subunit of ribulose-1,5-bisphosphate carboxylase from Oenothera is described. The peptides were characterized first by ion exchange chromatography and then by thin layer chromatography in a number of solvent systems, using ninhydrin and amino acid specific stains for detection. A comparison of the maps from different Oenothera species revealed considerable similarity between the species but with a small number of differences. The results are discussed in relation to the different chloroplast genomes present in Oenothera.

The cyanogen bromide fragments of the large subunit of ribulosebisphosphate carboxylase from barley

“…This was not the case for the cadmium ninhydrin orange peptide. Since peptides having N-terminal serine stain orange with cadmium ninhydrin (17), this peptide could be at the N-terminus of the fragment, which is in agreement with the Nterminal sequence (29) containing two histidine residues proximal to the first lysine at position 14.…”

“…A new aliquot of reference mixture was separated in the second dimension along the edge opposite to the sample application point. Subsequently, amino acid specific staining procedures were employed as described by HOLDF.R (17).…”

“…The derivative of threonine was observed only in one out of five experiments and that of serine in three out of five, probably due to low yields of cleavage of Met-Thr and Met-Ser peptide bonds (39) or to partial destruction during acid hydrolysis (24). The absence of small peptides, acylamino acids and amino acids in the fragments mixture obtained by CNBr-cleavage was ascertained by thin layer chromatograms stained with the C 12/starch/KI reagent (17).…”

“…A comparison of corresponding cyanogen bromide fragments from barley and spinach have revealed four amino acid differences among 37 analysed residues (29,42). Evidence that the gene for the large subunit in the chloroplast DNA is not completely conserved has also been obtained by chymotryptic peptide mapping of the large subunit from 9 genotypes of the genus Oenothera (17). The comparison included five different plastome types defined by their incompatibility relationships with nuclear genomes in interspecific crossings.…”

Peptide mapping of the ribulose bisphosphate carboxylase small subunit from the somatic hybrid of tomato and potato

“…Three 5 nmole samples from each of the three digests were chromatographed. Subsequently, the three sets of chromatograms were stained with cadmium ninhydrin, fluorescamine and with the amino acid specific stains as described by HOLDER (7,8) and by POUt,SEN (16). Thus, two plates from each set were stained with Cd-ninhydrin and with the tyrosine stain.…”