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ATPase activity and ATP/ADP-induced conformational change in the soluble domain of the bacterial protein translocator HlyB


, : ATPase activity and ATP/ADP-induced conformational change in the soluble domain of the bacterial protein translocator HlyB. Molecular Microbiology. 8(6): 1163-1175

The haemolysin exporter HlyB and its homologues are central to the unconventional signal-peptide-independent secretion of toxins, proteases and nodulation proteins by bacteria. HlyB is a member of the ATP-binding cassette (ABC) or traffic ATPase superfamily, and resembles closely in structure and function mammalian exporters such as the multidrug-resistance P-glycoprotein, combining both integral membrane and cytosolic domains. Overproduction of the HlyB cytoplasmic domain as a C-terminal peptide fused to glutathione S-transferase allowed the direct affinity purification and concentration of 30-50 mg ml-1 of soluble protein (GST-Bctp) in an apparently dimeric form possessing both transferase and ATPase activity. GST-Bctp bound to ADP-agarose and was eluted specifically by ATP and ADP, affinity behaviour which was confirmed in both the full-length HlyB and the unfused HlyB cytoplasmic domain synthesized in vitro. The stoichiometry of binding to MgATP and MgADP was close to equimolar and both ligands induced substantial conformational change in the protein. Mg-2+-dependent ATPase activity of GST-Bctp (V-max 1 mu-mol min-1 mg-1, K-m 0.2 mM) was comparable with the activity of the bacterial importer MalK and human P-glycoprotein reconstituted into proteoliposomes, and over an order of magnitude higher than in vitro measurements of disaggregated MalK purified from inclusion bodies. Activity was unaffected by inhibitors of F- and V-type ATPases, non-hydrolysable ATP analogues, or translocation substrate, but was severely inhibited by inhibitors of E1E2 (P-type) ATPases, and the acidic phospholipid phosphatidyl glycerol.

US$19.90

PMID: 8361361

DOI: 10.1111/j.1365-2958.1993.tb01661.x


Other references

Takata, K.U.da, K.K.izumi, A., 1995: Geographic variation in modulus of elasticity of tree trunks among Japanese larch planted in Hokkaido. Ecology and management of larix forests: a look ahead: proceedings of an international symposium Whitefish Montana USA October 5-9-1992 504

Tabb, D.L.; Smith, L.L.; Breci, L.A.; Wysocki, V.H.; Lin, D.; Yates, J.R., 2003: Statistical characterization of ion trap tandem mass spectra from doubly charged tryptic peptides. Collision-induced dissociation (CID) is a common ion activation technique used to energize mass-selected peptide ions during tandem mass spectrometry. Characteristic fragment ions form from the cleavage of amide bonds within a peptide undergoing C...

Jenkins, A.; Harrison, N.L.; Houamed, K.M., 1999: Interactions of volatile anesthetics with recombinant mammalian brain SK2 potassium channels. Anesthesiology (Hagerstown) 91(3A): A803, Sept

Krstic, Nadezda, 2002: Specific developmental disabilities Detection and interventions. While the majority of issues related to specific developmental disabilities (SDD) may be timely dealt with on the level of child's natural surrounding, especially through school-itself, in our community they have been, with sporadic exception...

Regitz-Zagrosek, V.; Seeland, U., 2011: Sex and gender differences in myocardial hypertrophy and heart failure. Cardiovascular disease is the most common cause of death in men and women worldwide. Men develop most, but not all, cardiovascular diseases at an earlier age while the number of affected women significantly increases with higher age. Heart failure...

Supe S.V.; Ganorkar P.L., 1982: Impact of operational research project wardha india. The impact of the Operational Research Project, Wardha is evaluated. The program made impact in terms of certain physical, economical, social and cultural development. The activities related to these objectives were studied. Implications are drawn...

Crockett, D.J., 1975: Diagnostic coding in a plastic surgery unit. British Journal of Plastic Surgery 28(4): 286-288

Wisniewski, E.W.; Albright, J.L.; Dillon, W.M.; Cunningham, M.D.; Taylor, R.W., 1977: Behavioral responses of heifers trained to enter the milking parlor by conditioning methods. 3 groups of heifers were allocated to one of 3 methods by which they were trained to enter the milking parlour: as soon as the door was opened; after 4 flashing red lights in the collecting yard were turned on; or after a buzzer was activated in t...

Cekan Z.; Robertson D.M.; Diczfalusy E., 1973: Calculation of radio immunoassay results on a wang programmable electronic calculator. Acta Endocrinologica Supplementum 177: 101

Moyer R.S., 1973: Comparing objects in memory evidence suggesting an internal psycho physics. Perception & Psychophysics 13(2): 180-184