title: Affinity chromatography as a means to study multienzyme complexes involved in murein synthesis.
abstract: The interaction of murein hydrolases and synthases was studied by affinity chromatography. The lytic transglycosylases Slt70 and MltB of E. coli were purified and covalently linked to CNBr-activated Sepharose. Membrane extracts were analyzed for proteins that interact with the immobilized murein hydrolases. Slt70-Sepharose was found to retain the PBPs 1b, 1c, 2, and 3. Likewise MltB-Sepharose enriched PBP 1b, 1c, and 3. Thus both lytic transglycosylases have an affinity for a transpeptidase, PBP2 and/or 3, as well as for the bifunctional transpeptidase/transglycosylase 1b. Interestingly, in addition, the poorly characterized PBP 1c interacts strongly with both Slt70 and MltB. It is speculated that the lytic transglycosylases assemble a multienzyme complex consisting of hydrolases and synthases, which is involved in growth of the stress-bearing murein sacculus.
authors.affiliation: Max-Planck-Institut für Entwicklungsbiologie, Tübingen, Germany.
authors.forename: M
authors.initials: M
authors.lastname: von Rechenberg
authors.forename: A
authors.initials: A
authors.lastname: Ursinus
authors.forename: J V
authors.initials: JV
authors.lastname: Höltje
mesh_terms: D002462:Cell Membrane / Q000201:enzymology; D002846:Chromatography, Affinity; D003488:Cyanogen Bromide; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D016695:Glycosyltransferases / Q000096:biosynthesis; D009097:Multienzyme Complexes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010457:Peptidoglycan / Q000096:biosynthesis*; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols; D012685:Sepharose
chemical_list: D009097:Multienzyme Complexes; D010457:Peptidoglycan; D011092:Polyethylene Glycols; D012685:Sepharose; D016695:Glycosyltransferases; C021815:murein transglycosylase; D003488:Cyanogen Bromide