Membrane-associated alkaline phosphatase from Bacillus licheniformis that requires detergent for solubilization: lactoperoxidase 125I localization and molecular weight determination

Spencer, Donald B.; Hansa, J G; Stuckmann, K V; Hulett, F. Marion

doi:10.1128/jb.150.2.826-834.1982

Cited by 14 publications

(4 citation statements)

References 26 publications

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“…This suggests a membrane-associated location for a-glucosidase, but all attempts to recover a-glucosidase activity from membranes have failed (G. Gammack & F. G. Priest, unpublished information). In common with alkaline phosphatase (Glynn et al, 1977;Spencer et al, 1982), the intracellular and extracellular a-glucosidases have essentially the same subunit molecular weights and could not be resolved by SDS-PAGE. This eliminates the involvement of a hydrophobic sequence containing a lipid moiety in the localization of these enzymes, as found in the membrane-bound penicillinase of this bacterium (Nielsen et al, 1981).…”

Section: Glucosementioning

confidence: 99%

Purification and Characterization of an Extracellular and a Cellular -Glucosidase from Bacillus licheniformis

Thirunavukkarasu

Priest

1984

Microbiology

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a-Glucosidase has been purified from culture fluid and from lysed cells of Bacillus licheniformis NCIB 6346. The enzymes from these two sources were virtually identical in molecular weight as judged by SDS-PAGE (63 000) and catalytic properties. The enzymes were unstable at high temperature and lost all activity after incubation at 60 "C for 10 min. Of the substrates examined, isomaltose gave maximal activity, followed by maltotriose, p-nitrophenyl a-D-glucopyranoside, sucrose and maltose. With isomaltose or maltotriose as substrate, transglucosylation activity was evident.

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Section: Glucosementioning

confidence: 99%

Purification and Characterization of an Extracellular and a Cellular -Glucosidase from Bacillus licheniformis

Thirunavukkarasu

Priest

1984

Microbiology

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“…The alkaline phosphatase of bacilli is salt extractable, especially by salts of magnesium (11,35). To gain insight into the possibility of changes in the nature of the binding of alkaline phosphatase to the membrane, we examined the extraction of the enzyme by different reagents.…”

Section: Resultsmentioning

confidence: 99%

Alkaline phosphatase secretion-negative mutant of Bacillus licheniformis 749/C

Kumar¹,

Ghosh²,

Ghosh³

1983

J Bacteriol

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An alkaline phosphatase secretion-blocked mutant of Bacillus licheniformis 749/C was isolated. This mutant had defects in the phoP and phoR regions of the chromosome. The selection procedure was based on the rationale that N-methyl-N'-nitro-N-nitrosoguanidine can induce mutations of closely linked multiple genes. The malate gene and the phoP and phoR genes are located at the 260-min position in the Bacillus subtilis chromosome; hence, the malate gene could be used as a marker for the mutation of the phoP and phoR regions of the chromosome. In a two-step selection procedure, strains defective in malate utilization were first selected with the cephalosporin C procedure. Second, these malate-defective strains were further screened in a dye medium to select strains with defects in alkaline phosphatase secretion. One stable mutant (B. licheniformis 749/C/NM105) had a total secretion block for alkaline phosphatase and had the following additional characteristics: (i) the amount of alkaline phosphatase synthesized was comparable to that in the wild type; (ii) the alkaline phosphatase was membrane bound; (iii) the mutant strain alkaline phosphatase, in contrast to that of the wild type, could not be extracted with MgCl2, although the amounts of protein extracted from each strain were comparable; (iv) the sodium dodecyl sulfate-polyacrylamide gel pattern of MgCl2-extracted proteins from the mutant strain was different from that of the wild-type proteins; (v) the mutant, unlike the wild type, could not use malate as a sole source of carbon; and (vi) the outside surface of the wall of the mutant cells contained an additional electron-dense layer that was not present on the wild-type cell wall surface.

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“…We investigated the binding of two secretory protein molecules, alkaline phosphatase and penicillinase, to the plasma membrane in bacilli. Penicillinase appears to bind to the membrane by hydrophobic interaction, whereas both hydrophobic and ionic interactions may be involved for alkaline phosphatase binding (24). Both enzymes are present in the membrane, but their distribution appears to be significantly different (8,9,11,26).…”

mentioning

confidence: 99%

Immunoelectron microscopic double labeling of alkaline phosphatase and penicillinase with colloidal gold in frozen thin sections of Bacillus licheniformis 749/C

Tinglu¹,

Ghosh²,

Ghosh³

1985

J Bacteriol

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The subcellular distribution of alkaline phosphatase and penicillinase was determined by double labeling frozen thin sections of Bacillus licheniformis 749/C with colloidal gold-immunoglobulin G (IgG). Antipenicillinase and anti-alkaline phosphatase antibodies were used to prepare complexes with 5-and 15-nm colloidal gold particles, respectively. The character of the labeling of membrane-bound alkaline phosphatase and penicillinase was different: the immunolabels for alkaline phosphatase (15-nm particles) were bound to a few sites at the inner surface of the plasma membrane, and the gold particles formed clusters of various sizes at the binding sites; the immunolabels for penicillinase (5-nm particles), on the other hand, were bound to the plasma membrane in a dispersed and random fashion. In the cytoplasm, immunolabels for both proteins were distributed randomly, and the character of their binding was similar. The labeling was specific: pretreating the frozen thin sections with different concentrations of anti-alkaline phosphatase or penicillinase blocked the binding of the immunolabel prepared with the same antibody. Binding could be fully blocked by pretreatment with 800 ,g of either antibody per ml.

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Membrane-associated alkaline phosphatase from Bacillus licheniformis that requires detergent for solubilization: lactoperoxidase 125I localization and molecular weight determination

Cited by 14 publications

References 26 publications

Purification and Characterization of an Extracellular and a Cellular -Glucosidase from Bacillus licheniformis

Purification and Characterization of an Extracellular and a Cellular -Glucosidase from Bacillus licheniformis

Alkaline phosphatase secretion-negative mutant of Bacillus licheniformis 749/C

Immunoelectron microscopic double labeling of alkaline phosphatase and penicillinase with colloidal gold in frozen thin sections of Bacillus licheniformis 749/C

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