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Titolo:
Cloning of the mspA gene encoding a porin from Mycobacterium smegmatis
Autore:
Niederweis, M; Ehrt, S; Heinz, C; Klocker, U; Karosi, S; Swiderek, KM; Riley, LW; Benz, R;
Indirizzi:
Univ Wurzburg, Lehrstuhl Biotechnol, Biozentrum, D-97074 Wurzburg, GermanyUniv Wurzburg Wurzburg Germany D-97074 entrum, D-97074 Wurzburg, Germany Univ Erlangen Nurnberg, Lehrstuhl Mikrobiol, D-91058 Erlangen, Germany Univ Erlangen Nurnberg Erlangen Germany D-91058 -91058 Erlangen, Germany Univ Calif Berkeley, Sch Publ Hlth, Div Publ Hlth Biol & Epidemiol, Berkeley, CA 94720 USA Univ Calif Berkeley Berkeley CA USA 94720 idemiol, Berkeley, CA 94720 USA ZymoGenet Inc, Seattle, WA 98102 USA ZymoGenet Inc Seattle WA USA 98102ZymoGenet Inc, Seattle, WA 98102 USA
Titolo Testata:
MOLECULAR MICROBIOLOGY
fascicolo: 5, volume: 33, anno: 1999,
pagine: 933 - 945
SICI:
0950-382X(199909)33:5<933:COTMGE>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
GRAM-NEGATIVE BACTERIA; OUTER-MEMBRANE PROTEIN; CELL-WALL PORIN; ESCHERICHIA-COLI; BIOPHYSICAL CHARACTERIZATION; SEQUENCE; TUBERCULOSIS; CHANNEL; ACID; PERMEABILITY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Benz, R Univ Wurzburg, Lehrstuhl Biotechnol, Biozentrum, Hubland, D-97074 Wurzburg, Germany Univ Wurzburg Hubland Wurzburg Germany D-97074 Wurzburg, Germany
Citazione:
M. Niederweis et al., "Cloning of the mspA gene encoding a porin from Mycobacterium smegmatis", MOL MICROB, 33(5), 1999, pp. 933-945

Abstract

Porins form channels in the mycolic acid layer of mycobacteria and therebycontrol access of hydrophilic molecules to the cell, We purified a 100 kDaprotein from Mycobacterium smegmatis and demonstrated its channel-forming activity by reconstitution in planar lipid bilayers. The mspA gene encodes a mature protein of 184 amino acids and an N-terminal signal sequence, MALDI mass spectrometry of the purified porin revealed a mass of 19 406 Da, in agreement with the predicted mass of mature MspA. Dissociation of the porinby boiling in 80% dimethyl sulphoxide yielded the MspA monomer, which did not farm channels any more, Escherichia coil cells expressing the mspA geneproduced the MspA monomer and a 100 kDa protein, which had the same channel-forming activity as whole-cell extracts of M. smegmatis with organic solvents. These proteins were specifically detected by a polyclonal antiserum that was raised to purified MspA of M. smegmatis, These results demonstrate that the mspA gene encodes a protein of M. smegmatis, which assembles to anextremely stable oligomer with high channel-forming activity, Database searches did not reveal significant similarities to any other known protein. Southern blots showed that the chromosomes of fast-growing mycobacterial species contain homologous sequences to mspA, whereas no hybridization could be detected with DNA from slow growing mycobacteria, These results suggest that MspA is the prototype of a new class of channel-forming proteins.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 13:11:05