Production and characterization of recombinant P1 adhesin essential for adhesion, gliding, and antigenic variation in the human pathogenic bacterium, Mycoplasma pneumoniae
| Autor: | Shigetarou Mori, Tsuyoshi Kenri, Yuuki Hayashi, Hisashi Kudo, Yuhei O Tahara, Munehito Arai, U Matsumoto, Yukio Furukawa, Makoto Miyata, Yoshito Kawakita, Keigo Shibayama |
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| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular 0301 basic medicine Circular dichroism Biophysics medicine.disease_cause Biochemistry Bacterial Adhesion law.invention 03 medical and health sciences 0302 clinical medicine X-Ray Diffraction law Scattering Small Angle medicine Humans Adhesins Bacterial Molecular Biology Escherichia coli Host cell surface chemistry.chemical_classification Chemistry Cell Biology Antigenic Variation Recombinant Proteins Amino acid Bacterial adhesin Attachment organelle Transmembrane domain 030104 developmental biology 030220 oncology & carcinogenesis Hydrodynamics Recombinant DNA |
| Zdroj: | Biochemical and Biophysical Research Communications. 508:1050-1055 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2018.11.132 |
| Popis: | Mycoplasma pneumoniae forms an attachment organelle at one cell pole, binds to the host cell surface, and glides via a unique mechanism. A 170-kDa protein, P1 adhesin, present on the organelle surface plays a critical role in the binding and gliding process. In this study, we obtained a recombinant P1 adhesin comprising 1476 amino acid residues, excluding the C-terminal domain of 109 amino acids that carried the transmembrane segment, that were fused to additional 17 amino acid residues carrying a hexa-histidine (6 × His) tag using an Escherichia coli expression system. The recombinant protein showed solubility, and chirality in circular dichroism (CD). The results of analytical gel filtration, ultracentrifugation, negative-staining electron microscopy, and small-angle X-ray scattering (SAXS) showed that the recombinant protein exists in a monomeric form with a uniformly folded structure. SAXS analysis suggested the presence of a compact and ellipsoidal structure rather than random or molten globule-like conformation. Structure model based on SAXS results fitted well with the corresponding structure obtained with cryo-electron tomography from a closely related species, M. genitalium. This recombinant protein may be useful for structural and functional studies as well as for the preparation of antibodies for medical applications. |
| Databáze: | OpenAIRE |
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