Phage lambda receptor protein from <i>Escherichia coli</i>

Sandermann, Heinrich; Bavoil, P; Nikaido, Hiroshi

doi:10.1016/0014-5793(78)80062-3

Cited by 13 publications

(5 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The Css-isoprenoid-alcohol kinase has previously been highly purified in butan-1-01 [6,7]. The lactose permease protein [lo] and the phage lambda receptor protein [21] of E. coli have been isolated by use of hexamethylphosphoric triamide. This aprotic organic solvent was particularly effective in the solubilization of membrane proteins [16], and it also dissolved bacteriorhodopsin in high yield (H. Sandermann, unpublished results).…”

Section: Test Of C~~-~soprenogalcohol Kinasementioning

confidence: 99%

Diglyceride Kinase from Escherichia coli

Bohnenberger

Sandermann

1979

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

The diglyceride kinase activity of membranes from Escherichia coli was extracted into acidic butan-1-01. The enzyme was purified in organic solvent by precipitation at -20 "C, chromatography on DEAE-cellulose and repeated chromatography on Sephadex LH-60. The final 1460-fold purified enzyme preparation gave a single protein band upon isoelectric focusing in the presence of Triton X-100 (PI, 4.0) and upon polyacrylamide-gel electrophoresis in the presence of sodium dodecylsulphate. The latter method as well as gel chromatography on Sephadex LH-60 indicated a molecular weight of about 15 400.The purified enzyme was devoid of lipid, and it required re-addition of lipid for activity. sn-1,2-Dipalmitate and ceramide were phosphorylated, whereas the Css-isoprenoid alcohol, ficaprenol, did not serve as a substrate under the same conditions. Conversely, the butanol-soluble Cssisoprenoid-alcohol kinase from Staphylococcus aureus did not phosphorylate sn-l,2-dipalmitate.

show abstract

Section: Test Of C~~-~soprenogalcohol Kinasementioning

confidence: 99%

Diglyceride Kinase from Escherichia coli

Bohnenberger

Sandermann

1979

European Journal of Biochemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…To examine the function of the lamB protein in vitro, we developed a purification scheme that utilized neither sodium dodecyl sulfate (NaDodSO4) nor the organic solvents used in earlier works (5)(6)(7)(8). These agents could alter the activities of the protein by partially denaturing it, so we wanted to employ milder treatments to extract and purify the protein.…”

mentioning

confidence: 99%

Specificity of diffusion channels produced by lambda phage receptor protein of Escherichia coli.

Luckey¹,

Nikaido²

1980

Proc. Natl. Acad. Sci. U.S.A.

254

183

View full text Add to dashboard Cite

The lamB protein, the receptor for phage X, was purified from the outer membrane of Escherichia coli K-12 by extraction with Triton X-100 and EDTA, chromatography on DEAE-Sephacel in Triton X-100, exchange of Triton for cholate by gel filtration, and chromatography on Sephacryl S-200 in cholate, NaCI, and EDTA. The purified protein appeared to exist as several oligomeric species. In an equilibrium retention assay with reconstituted vesicles containing phospholipids and lipopolysaccharide, the lamB protein conferred permeability for disaccharides. In a liposome swelling assay designed to measure rates of diffusion, the lamB protein conferred permeability to phospholipid liposomes for a variety of substrates. The rates obtained indicate the permeation facilitated by the lamB protein is specific, discriminating among substrates by both size and configuration. For example, maltose diffused into liposomes 40 times faster than sucrose, about 8 times faster than cellobiose, and about 12 times faster than maltoheptaose. The results suggest that the lamB protein forms a transmembrane channel containing a site (or sites) that loosely interacts with the solutes.The outer membrane of Gram-negative bacteria has several pathways for the translocation of small molecules (reviewed in ref. 1). The passage of most small, hydrophilic molecules is accomplished by porins, which form nonspecific water-filled pores. Other outer membrane proteins are involved in the transport of specific nutrients, but the mechanism of the specific transport is not understood.The A phage receptor protein, product of the lamB gene (2), is required in Escherichia coli for the transport of maltose at micromolar concentrations and of maltodextrins of higher molecular weight (3). Because direct binding studies have failed to detect an affinity of lamB protein for maltose (4), the hypothesis that it also forms a fairly nonspecific pore has been attractive. Conductivity studies on black lipid membranes containing the lamB protein give evidence for a pore of fairly large diameter (5). Furthermore, while the current work was in progress, a paper by Nakae (6) appeared in which he claimed that membranes reconstituted from phospholipids, lipopolysaccharides, and lamB protein are nonspecifically permeable to all sugars tested except stachyose. In order to reconcile this apparent lack of specificity in reconstituted systems with the specificity for maltose and maltodextrins observed in intact cells, it has been proposed that the lamB protein produces a totally nonspecific channel, and that the specificity is derived entirely from the interaction of periplasmic maltose-binding protein with the lamB protein (7).To examine the function of the lamB protein in vitro, we developed a purification scheme that utilized neither sodium dodecyl sulfate (NaDodSO4) nor the organic solvents used in earlier works (5-8). These agents could alter the activities of the protein by partially denaturing it, so we wanted to employ milder treatments to extract and purify the prot...

show abstract

“…Protein was determined in the presence of SDS [3]. SDS gel electrophoresis was performed on 13% (w/v) polyacrylamide slab gels [6], samples being dissolved in the presenceof 2%(w/v)SDS(lOmin, lOO"C).sn-1.2. Dipalmitate and bovine heart cardiolipin were obtained from Fluka (Neu-Ulm) and Sigma (St Louis), respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Previous studies have shown a drastic influence of divalent ions upon total solubilization and upon the nature of polypeptides dissolved by HMPT [5,6].…”

Section: Solubiliza Tion Proceduresmentioning

confidence: 97%

“…Precipitation with ethanol and gel permeation chromatography on Sephacryl were developed as methods for protein purification in HMPT. The following defined membrane proteins have so far been solubilized and purified in HMPT, the Escherichia coli phage lambda receptor protein [6], bacterioopsin [7], the pig kidney Na+,K+-ATPase subunits [S. Maier, H. S. unpublished] and certain erythrocyte and brain membrane proteins [R. Schmitt, S. Maier, H. S., unpublished].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation