title: Characterization of high molecular weights of complexes and polymers of cytoplasmic proteins in Escherichia coli.
abstract: To search for filamentous polymers of cytoplasmic proteins of Escherichia coli, high molecular weights (> 670 kDa) of protein complexes of cell extracts were fractionated by gel filtration and ion-exchange column chromatography. Proteins of 100, 77 and 52 kDa were co-purified. The 100- and 52-kDa proteins were identified to be pyruvate dehydrogenase and lipoamide dehydrogenase, respectively, by determining the N-terminal amino acid sequences. Experimental results indicate that the 77-kDa protein is identical to dihydrolipoamide acyltransferase. The 100-kDa protein was found to be identical to the 100-kDa protein described by Tomioka (1991), and was related to the formation of filaments and sheets in the presence of 100 mM KCl. However, neither long filaments nor sheets were observed in our sample containing these enzymes, which was not consistent with Tomioka's conclusion. Another 100-kDa protein which forms spirosome-like particles was purified and identified to be alcohol dehydrogenase based on the N-terminal sequence.
authors.affiliation: Department of Molecular Cell Biology, Kumamoto University School of Medicine, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Imamura
authors.forename: H
authors.initials: H
authors.lastname: Niki
authors.forename: M
authors.initials: M
authors.lastname: Kitaoka
authors.forename: K
authors.initials: K
authors.lastname: Yamanaka
authors.forename: T
authors.initials: T
authors.lastname: Ogura
authors.forename: S
authors.initials: S
authors.lastname: Hiraga
mesh_terms: D000217:Acyltransferases / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000302:isolation & purification*; D003593:Cytoplasm / Q000737:chemistry*; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry*; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008970:Molecular Weight; D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D011768:Pyruvate Dehydrogenase Complex; D008058:Dihydrolipoamide Dehydrogenase; D000217:Acyltransferases; C033588:dihydrolipoamide acyltransferase