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

Poulsen, Carsten Stig

doi:10.1007/bf02906296

Cited by 20 publications

(19 citation statements)

References 35 publications

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“…The identity of the released amino acids was confirmed by dansylation as. described by POULSEN (12). Weak spots of glutamic acid and aspartic acid were found after hydrolysis for 4 hours in addition to the residues shown in the figure.…”

Section: Discussionmentioning

confidence: 77%

Purification of hordein polypeptides by column chromatography using volatile solvents

Schmitt

1979

Carlsberg Res. Commun.

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Section: Discussionmentioning

confidence: 77%

Purification of hordein polypeptides by column chromatography using volatile solvents

Schmitt

1979

Carlsberg Res. Commun.

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“…This used a comparison to the available amino acid sequence data from the barley and the spinach large subunit (44,45). Only two long stretches of sequence (nucleotides 451-824 and 1190-1575) without appropriate restriction endonuclease sites for 32p-labelling with 5' polynucleotide kinase or with the KLENOW fragment of DNA polymerase caused some difficulty.…”

Section: Determination Of the Large Subunit Primary Structurementioning

confidence: 99%

“…The LS was prepared in the usual way and cleaved with CNBr. The fragments were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (44) and the N-terminal amino acid residues determined by the dansylation procedure. The Nterminal fragments from both species had the same size and the dansylation procedure failed to yield an N-terminal amino acid, most likely due to N-terminal blocking.…”

Section: Determination Of the Large Subunit Primary Structurementioning

confidence: 99%

“…The purpose of the present paper is to relate the enzymological data obtained by active site modifications (13,26,27,38,42,50,52,55)of the catalytically active large subunit of the enzyme to its primary structure as determined by amino acid sequencing (44,45) and nucleotide sequencing of the gene for this polypeptide in maize (34) and spinach (63).…”

Section: Introductionmentioning

confidence: 99%

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Comments on the structure and function of the large subunit of the enzyme ribulose bisphosphate carboxylase-oxygenase

Poulsen

1981

Carlsberg Res. Commun.

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The complete amino acid sequence of the large subunit (LS) of ribulose bisphosphate carboxylase has been determined for maize through nucleotide sequencing of the chloroplast gone encoding this polypeptide (34). Previously published amino acid sequences from the large subunits of barley and spinach as well as new evidence for the amino acid sequence of these two polypeptides are aligned with the complete maize sequence. The complete spinach amino acid sequence is also presented according to the sequence of the spinach gene (63). The number of amino acids in the 475-residue LS primary structure for which there is direct or circumstantial evidence from barley, is 278. Amino acid replacements are encountered in seven positions in comparison with the maize sequence and in 20 positions when compared to the spinach LS. From affinity labelling studies ( 13, 26) active site residues, nominally lysines and cysteines, have been deduced and located in the total maize and spinach LS sequences, thus being available for further study of structure-function relationships. The complete amino acid sequence and knowledge about catalytic site residues also allows direct comparison with primary structures of other enzymes of the same class, the lyases. Particular attention is devoted to aldolase, an enzyme also functioning in carbohydrate metabolism. Comparison with the 361-residue aldolase of glycolysis in rabbit muscle cells suggests homology in 67 positions centered around catalytic site lysine-175 of RuBPCase LS and catalytic site lysine-227 of aldolase. It is discussed how this relationship may throw light on the reaction mechanism of the RuBP carboxylation.Abbreviations: BBBP = D-3-bromo-l,4-dihydroxy-butanone-l,4-bisphosphate; CA = carbonic anhydrase; CABP = D-2-carboxyarabinitoI-1,5-bisphosphate; DHAP = dihydroxyacetonephosphate; FBP = Dfructose-l,6-bisphosphate; FBPald = fructose-l,6-bisphosphate aldolase; 1,3GABP = glyceric acid-l,3-bisphosphate; G3P = glyceraldehyde-3-phosphate; G3PDH = (NADP)-glyceraldehyde-3-phosphate dehydrogenase; G3Piso = G3P ketol-isomerase; LS = large subunit; NBA-EAP = N-bromoacetyl-ethanolamine phosphate; 3PGA = 3-phosphoglycerate; 3PGAkin = 3-phosphoglycerate kinase; 6PGIuA = D-6-phosphogluconate; PLP = pyridoxal-5"-phosphate; RuBP = D-ribulose-l,5-bisphosphate; RuBPCase = RuBP carboxylase; SS = small subunit.

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“…Hydrolysis and amino acid analysis of small subunit samples was carried out as previously described for the large subunit of RuBPCase from barley (16). In all cases about 25 lag of small subunit was hydrolyzed.…”

Section: Amino Acid Analysismentioning

confidence: 99%

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

Poulsen

Porath

Sacristán

et al. 1980

Carlsberg Res. Commun.

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K e y w o r d s : F r a c t i o n 1 protein, isoelectric focusing, c h y m o t r y p t i c peptides, tryptic peptides, protoplast fusionFive somatic hybrids of potato and tomato obtained by protoplast fusion were identified using the isoelectric focusing patterns of ribulose bisphosphate carboxylase as phenotypic markers of the nuclear and chloroplast genomes. Together with the previously analyzed hybrids (14) this brings the number of verified somatic hybrids between the two species to nine. The products of both tomato and potato nuclear genomes are present in the ribulose bisphosphate carboxylase oligomer from all nine hybrids. Four of the hybrids contain the tomato large subunit and hence the tomato chloroplast DNA is functional, whereas in five hybrids the potato ribulose bisphosphate carboxylase large subunit and thereby the potato chloroplast DNA is present. The former are designated topatoes and the latter pomatoes (I 1).Mapping of chymotryptic peptides of the small subunit of tomato and potato revealed a tyrosine containing peptide with species specific mobility. Peptide maps from an analyzed somatic hybrid contained both the tyrosine peptide specific for potato and that specific for tomato. Maps of tryptic peptides of the small subunit of tomato differed from those of the potato in 2 peptides. The tryptic peptide maps of one analyzed hybrid contained the two peptides unique to potato as well as the two peptides unique to tomato. The tomato-potato hybrids are aneuploid with chromosome numbers ranging from 48 to 74, the expected allotetraploid number being 48 and the expected allohexaploid number 72.Abbreviations: RuBPCase = ribulose-l,5-bisphosphate carboxylase; DTT --dithiothreitol; SDS = sodium dodecyl sulphate.

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

Cited by 20 publications

References 35 publications

Purification of hordein polypeptides by column chromatography using volatile solvents

Purification of hordein polypeptides by column chromatography using volatile solvents

Comments on the structure and function of the large subunit of the enzyme ribulose bisphosphate carboxylase-oxygenase

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

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