Mycoplasma pneumoniae adhesin localized to tip structure by monoclonal antibody

Feldner, J; Göbel, Ulf B.; Bredt, W.

doi:10.1038/298765a0

Cited by 158 publications

(120 citation statements)

References 18 publications

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“…The adhesion molecule of M. pneumoniae, designated P1, is a 170-kDa protein containing at least three transmembrane domains with most of the protein outside of the cell (26). P1 protein is located mainly at the attachment organelle with some accessory cytadherence proteins (1,8,14,33). The mutant lacking P1 protein cannot bind to glass (26) and consequently cannot glide.…”

Section: Discussionmentioning

confidence: 99%

Force and Velocity of Mycoplasma mobile Gliding

2002

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The effects of temperature and force on the gliding speed of Mycoplasma mobile were examined. Gliding speed increased linearly as a function of temperature from 0.46 m/s at 11.5°C to 4.0 m/s at 36.5°C. A polystyrene bead was attached to the tail of M. mobile using a polyclonal antibody raised against whole M. mobile cells. Cells attached to beads glided at the same speed as cells without beads. When liquid flow was applied in a flow chamber, cells reoriented and moved upstream with reduced speeds. Forces generated by cells at various gliding speeds were calculated by multiplying their estimated frictional drag coefficients with their velocities relative to the liquid. The gliding speed decreased linearly with force. At zero speed, the force measurements extrapolated to 26 pN at 22.5 and 27.5°C. At zero force, the speed extrapolated to 2.3 and 3.3 m/s at 22.5 and 27.5°C, respectively-the same speeds as those observed for free gliding cells. Cells attached to beads were also trapped by an optical tweezer, and the stall force was measured to be 26 to 28 pN (17.5 to 27.5°C). The gliding speed depended on temperature, but the maximum force did not, suggesting that the mechanism is composed of at least two steps, one that generates force and another that allows displacement. Other implications of these results are discussed.Mycoplasmas are parasitic bacteria with a small genome and no peptidoglycan layer (27). Several mycoplasma species have a distinct cell polarity characterized by a protruding membrane extension, the attachment organelle (27). They are able to attach to and glide on glass, plastic, and eukaryotic cell surfaces, always moving in the direction of the organelle (19). The gliding mechanism is unknown. Mycoplasmas do not have any appendages such as flagella or pili (19) or any genes obviously related to motility, including motor proteins such as myosin or kinesin (7, 10, 13). However, a transmembrane protein associated with a cytoskeleton-like structure has been shown to be necessary for glass binding in Mycoplasma pneumoniae (22).Mycoplasma mobile, isolated from a fish gill organ, has a conical cell structure, with the attachment organelle at the apex of the cone. It glides up to four cell lengths per second (2.5 m/s) in the direction of the apex, which is designated as a head-like structure (30). However, little is known about force generation (28,29). In this study, to characterize its gliding motility, we attached a bead to the tail end of M. mobile and measured the gliding force as a function of speed using viscous flow and an optical tweezer. MATERIALS AND METHODSCultivation. M. mobile strain 163K (ATCC 43663) was grown as described previously (25).Measurements of gliding speed. M. mobile cells in a culture at an optical density at 600 nm of 0.03 to 0.1 were collected by centrifugation at 10,000 ϫ g for 4 min at room temperature and resuspended in a fivefold-smaller volume of medium. A 5-l aliquot was sealed between a coverslip and a slide within a thin ring of Apiezon M grease (Apiezon Products, L...

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Section: Discussionmentioning

confidence: 99%

Force and Velocity of Mycoplasma mobile Gliding

2002

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“…The protein with the higher molecular mass, estimated at 170-190 kDa, was named P1 (Hu et al, 1977). Subsequent studies established that protein PI is the major adhesin of M. pneumoniae (Hu et al, 1982;Baseman et al, 1982;Feldner et al, 1982). M .…”

Section: The P1 Operon Genes and Proteinsmentioning

confidence: 99%

Mycoplasma adhesion

Razin¹,

Jacobs²

1992

Journal of General Microbiology

232

222

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“…A critical step in bacterial colonization of the host cells is the specific adhesion to host cell receptors, mediated by bacterial adhesins. In M. pneumoniae, the P1 adhesin (3,17,25,26) and the adhesin-related 30-kDa protein (4, 10) have been identified. Both proteins are located mainly in a tip structure that functions as the attachment organelle of the bacterium.…”

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The proline-rich P65 protein of Mycoplasma pneumoniae is a component of the Triton X-100-insoluble fraction and exhibits size polymorphism in the strains M129 and FH

et al. 1995

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Previously, we described the identification of a novel Mycoplasma pneumoniae M129 protein, named P65 because of its apparent molecular mass of 65 kDa estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (T. Proft and R. Herrmann, Mol. Microbiol. 13:337-348, 1994). DNA sequence analysis of the P65 open reading frame (orfp65), however, revealed an ORF encoding a protein with a molecular weight of 47,034. This discrepancy can be explained by the unusual amino acid composition of this protein. According to the deduced amino acid sequence, the N-terminal half of P65 contains several penta-and hexapeptides (DPNAY and DPNQAY) forming a proline-rich acidic domain. Secondary-structure predictions indicated ␤-sheets and turns within that region, suggesting an extended and rigid conformation. Near the C terminus of P65 the tripeptide Arg-Gly-Asp (RGD) was found. This motif is known to play an important role in binding of extracellular matrix proteins to integrins. P65 could be located exclusively to the Triton X-100-insoluble cell fraction. The results of immunofluorescence microscopy and of immunoadsorption experiments indicated that P65 carries surface-exposed regions. Mild treatment of whole cells with proteases resulted in cleavage of a limited amount of P65 molecules, suggesting either that only a small percentage of P65 molecules are exposed on the surface or that protease cleavage is hampered by a compact protein conformation or by binding of an unknown component to P65. P65 exhibits size polymorphism in M. pneumoniae M129 and FH. This is caused by an intragenetic duplication of a 54-bp sequence within the FH orfp65. As a consequence, the number of DPNAY pentapeptides increased from 9 to 12 repeats in the FH strain.Mycoplasma pneumoniae is an extracellular pathogen of the human respiratory tract (26, 46) causing histopathological changes of lung epithelial cells, usually in older children and young adults (18). A critical step in bacterial colonization of the host cells is the specific adhesion to host cell receptors, mediated by bacterial adhesins. In M. pneumoniae, the P1 adhesin (3,17,25,26) and the adhesin-related 30-kDa protein (4, 10) have been identified. Both proteins are located mainly in a tip structure that functions as the attachment organelle of the bacterium. It could be demonstrated, by nearestneighbor analysis with a hydrophilic chemical cross-linker, that the product of open reading frame 6 (ORF6) of the P1 operon (28), a 40-kDa protein and 90-kDa protein (9, 34, 50) are located in close proximity to the P1 adhesin on the cell surface (35).Scanning and transmission electron microscope analyses of M. pneumoniae cells grown on grids and pretreated with Triton X-100 revealed a rodlike tip structure and a network of filamentous strands (22,41). Krause and coworkers identified a set of high-molecular-weight proteins (HMW1 to HMW5) which play an important role in cytadherence (30,31,(51)(52)(53) and appear to be involved in formation of a cytoskeletonlike structure (38). These proteins, ...

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Mycoplasma pneumoniae adhesin localized to tip structure by monoclonal antibody

Cited by 158 publications

References 18 publications

Force and Velocity of Mycoplasma mobile Gliding

Force and Velocity of Mycoplasma mobile Gliding

Mycoplasma adhesion

The proline-rich P65 protein of Mycoplasma pneumoniae is a component of the Triton X-100-insoluble fraction and exhibits size polymorphism in the strains M129 and FH

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