A mass-spectrometric sequence study of the enzyme ribitol dehydrogenase from <i>Klebsiella aerogenes</i>

Morris, Howard R.; Williams, Dudley H.; Midwinter, Graeme G.; Hartley, Brian

doi:10.1042/bj1410701

Cited by 76 publications

(13 citation statements)

References 11 publications

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“…Fortunately the vast majority of ambiguities were resolved by this approach. The peptide mixtures isolated from the thermolysin digest, in the present work, gave rise to the same difficulties as those described by Morris et al [4]. Amino acids common in the N-terminal portions of such peptides have molecular weights in a numerical relationship which leads to ambiguities in sequence assignments.…”

Section: ~~O -~~~-~~~-~~~: -~~~-~~~-~~~-~~~-~~~-~~~-~-A L a -A S~-a~-supporting

confidence: 67%

“…The fragmentations of acetyl-permethyl-peptides are well documented [4,12]. Except where stated below, the peptides studied here behaved in a similar manner, and the amount of sequence information derived is summarised in Table 1.…”

Section: Resultsmentioning

confidence: 74%

“…Although access to information from that study was available, all mass spectra have been interpreted independently. The strategy employed in this study, in particular the use of non-specific proteinases, although proposed by Morris et al [4] was not used exclusively in their work. We have found that the use of non-specific enzymes followed by gel filtration and paper electrophoresis tends to produce mixtures of peptides of similar sizes and hence similar thermal volatilities.…”

Section: ~~O -~~~-~~~-~~~: -~~~-~~~-~~~-~~~-~~~-~~~-~-A L a -A S~-a~-mentioning

confidence: 99%

“…This approach has also been applied to large proteins and is illustrated by the study of ribitol dehydrogenase [4].…”

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confidence: 99%

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Mass Spectrometric Sequence Studies of a Superoxide Dismutase from Bacillus stearothermophilus

Auffret

Blake²,

Williams

1981

European Journal of Biochemistry

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A partial sequence of the manganese-containing superoxide dismutase from Bacillus stearothermoplzilus has been determined by mass spectrometry. A new fragmentation reaction at N-terminal proline is described and an 'in-chain' fragmentation, also occurring at proline, has been observed. There is some evidence that the latter cleavage may also occur at other residues. Some differences from the classically determined sequence are reported, and the validity of these observations is discussed.Currently there are two strategies in use for the mass spectrometric determination of protein sequences. One requires the generation of small peptides (of two to five residues) and their derivatisation to 0-trimethylsilylated trifluoroacetyl-poly(amino alcohols). Separation and identification of these derivatives is accomplished by linked gas chromatography and mass spectrometry. This strategy has been successfully used in the sequencing of several proteins up to 250 amino acid residues [I -31.The other method involves the analysis of mixtures of acetyl, N-permethyl and 0-permethyl peptides, usually generated by the use of proteinases familar in conventional sequence work. This approach has also been applied to large proteins and is illustrated by the study of ribitol dehydrogenase [4].

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Section: ~~O -~~~-~~~-~~~: -~~~-~~~-~~~-~~~-~~~-~~~-~-A L a -A S~-a~-supporting

confidence: 67%

Section: Resultsmentioning

confidence: 74%

Section: ~~O -~~~-~~~-~~~: -~~~-~~~-~~~-~~~-~~~-~~~-~-A L a -A S~-a~-mentioning

confidence: 99%

“…This approach has also been applied to large proteins and is illustrated by the study of ribitol dehydrogenase [4].…”

mentioning

confidence: 99%

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Mass Spectrometric Sequence Studies of a Superoxide Dismutase from Bacillus stearothermophilus

Auffret

Blake²,

Williams

1981

European Journal of Biochemistry

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“…The ORF4 product is similar to a group of oxidoreductases ( Fig. 3), including the protochlorophyllide reductase (Pcr) of barley (33), the glucose dehydrogenase of Bacillus megateium (19), the ribitol dehydrogenase of Kgebsiella aerogenes (28), and the NodG protein of Rhizobium meliloti (which is homologous to ,B-ketoacyl reductase [34]). The similarity among these oxidoreductases was stronger at the N termini.…”

Section: Methodsmentioning

confidence: 99%

Discovery and characterization of a new transposable element, Tn4811, in Streptomyces lividans 66

Chen¹,

Yu²,

Chung³

et al. 1992

J Bacteriol

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Transposition of a new 5.4-kb transposon, Tn4811, of Streptomyces lividans to the melC operon of Streptomyces antibioticus on plasmid pU702 was discovered. The nucleotide sequence of this copy of Tn4811, which contained an imperfect (9 of 11 bp) terminal inverted repeat, five putative Streptomyces coding sequences for an oxidoreductase and its transcription regulator, and three transposition-related proteins, was determined. SLP-strains of S. lividans contained one copy (A) of Tn4811, while SLP2+ strains contained an additional copy (B) on the SLP2 plasmid. The nucleotide sequences at three insertion junctions of Tn4811 were determined. Copy B lacked 41 bp from the left end. At the other five junctions the duplication of a putative 3-bp target sequence (TGA) was observed. A sequence of less than 3 kb homologous to Tn4811 was present in S. antibioticus. DNA homologous to Tn4811 was not detected in 14 other Streptomyces species.Structural instability of genomic DNA is a widespread feature of gram-positive, filamentous, soil bacteria of the genus Streptomyces (32). Many genetic traits undergo spontaneous loss in laboratory cultures at frequencies of 10' to 10-2. Exposure to certain stresses such as UV irradiation, DNA intercalating agents, cold temperature, or protoplasting and regeneration increases the frequency of these instabilities. The mutations involved in the instabilities are attributed to large deletions of chromosomal DNA, which are frequently accompanied by tandem amplifications of particular sequences nearby (32). The genetic principle(s) underlying the structural instability is not clear. Possible mechanisms involved are homologous recombination, site-specific recombination, and transposition. Homologous recombination systems in Streptomyces species and their contribution to genetic instability have received little study. Tsai and Chen (41) isolated a rec mutant defective in intraplasmid recombination but not in chromosomal recombination (25). Chou and Chen (11), in their investigation of an unstable arg gene, found no effect of this rec mutation on its instability. Site-specific recombination and transposition, although representing forms of fluidity of genomic DNA, have not been implicated in the instability of other DNA sequences (8).About one-third of Streptomyces species can produce melanin pigment (44). The melanin (melC) operon of Streptomyces antibioticus has been cloned (24), sequenced (3), and widely used in many recombinant vectors. The melC operon in the Streptomyces species examined is genetically unstable, undergoing spontaneous deletions at relatively high frequencies-about 10-' (32). Similar to those observed in other unstable genes in Streptomyces species, the deletions of melC were frequently accompanied by extensive tandem amplifications of specific sequences (18,32).

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Sequenzanalyse von Polypeptiden und Proteinen mit der kombinierten Gaschromatographie‐Massenspektrometrie

Nau

1976

Angewandte Chemie

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Bei einer neuen Methode zur Bestimmung der Aminosiiure-Sequenz wird das Polypeptid zunachst durch Partialhydrolyse in ein komplexes Gemisch von Oligopeptiden zerlegt, das durch Derivatisierung fliichtiger gemacht und schlieDlich mit der kombinierten Gaschromatographie-Massenspektrometrie analysiert wird. Aus den dabei identifizierten Oligopeptiden ermittelt ein Computer die Sequenz des Polypeptids. Bisher gelang auf diese Weise die Sequenzanalyse von Polypeptiden mit bis zu etwa 40 Aminosiiuren. Die Vor-und Nachteile dieser neuen Methode gegeniiber dem Edman-Abbau-Verfahren rnit seinem stufenweisen Vorgehen werden abgewogen. Da beide Methoden von prinzipiell unterschiedlichen Voraussetzungen ausgehen, konnen sie sich erganzen.

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A mass-spectrometric sequence study of the enzyme ribitol dehydrogenase from Klebsiella aerogenes

Cited by 76 publications

References 11 publications

Mass Spectrometric Sequence Studies of a Superoxide Dismutase from Bacillus stearothermophilus

Mass Spectrometric Sequence Studies of a Superoxide Dismutase from Bacillus stearothermophilus

Discovery and characterization of a new transposable element, Tn4811, in Streptomyces lividans 66

Sequenzanalyse von Polypeptiden und Proteinen mit der kombinierten Gaschromatographie‐Massenspektrometrie

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