Polypeptide cartography of Spiroplasma taiwanense.
Spiroplasma taiwanense is the first member of the Class Mollicutes to be subjected to polypeptide cartography using computerized image analysis. The small genome size characteristic of this group was shown to code for low numbers of polypeptides when compared to other bacterial species. Silver-stained two-dimensional electrophoresis gels, following separation by either isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (ISO-DALT) or nonequilibrium pH gradient electrophoresis (NEPHGE), were used to create databases from 10 and 6 gels, respectively, for each technique and produced, respectively, 263 and 287 replicated spots. Polypeptides were mapped with respect to molecular mass and glyceraldehyde-3-phosphate dehydrogenase carbamylation standards. Of interest was the unexpectedly high percentage (50.2%) of the total normalised optical intensity associated with all 263 spots detected by ISO-DALT electrophoresis, having been contributed by just 29 dominant protein spots. These 29 polypeptides are to be given priority in microsequencing and microanalysis aimed at their identification.
Humphery-Smith
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Départment de Microbiologie et Santé Publique, Faculté de Médecine, Brest.
Guyonnet
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Chastel
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C
D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis; D007525:Isoelectric Focusing / Q000379:methods; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification*; D012015:Reference Standards; D015203:Reproducibility of Results; D013149:Spiroplasma / Q000737:chemistry* / Q000254:growth & development
D001426:Bacterial Proteins; D010455:Peptides; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases