Study of the protein complex, pore diameter, and pore-forming activity of the Borrelia burgdorferi P13 porin

P13 is one of the major outer membrane proteins of Borrelia burgdorferi. Previous studies described P13 as a porin. In the present study some structure and function aspects of P13 were studied. P13 showed according to lipid bilayer studies a channel-forming activity of 0.6 nanosiemens in 1 M KCl. Single channel and selectivity measurements demonstrated that P13 had no preference for either cations or anions and showed no voltage-gating up to +/-100 mV. Blue native polyacrylamide gel electrophoresis was used to isolate and characterize the P13 protein complex in its native state. The complex had a high molecular mass of about 300 kDa and was only composed of P13 monomers. The channel size was investigated using non-electrolytes revealing an apparent diameter of about 1.4 nm with a 400-Da molecular mass cut-off. Multichannel titrations with different substrates reinforced the idea that P13 forms a general diffusion channel. The identity of P13 within the complex was confirmed by second dimension SDS-PAGE, Western blotting, mass spectrometry, and the use of a p13 deletion mutant strain. The results suggested that P13 is the protein responsible for the 0.6-nanosiemens pore-forming activity in the outer membrane of B. burgdorferi.

Thein, Marcus (författare)
Barbot, Mariam (författare)
Sans-Serramitjana, Eulalia (författare)
Bonde, MariUmeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten)(Swepub:umu)maiboe01 (författare)
Mentele, Reinhard (författare)
Lottspeich, Friedrich (författare)
Bergström, SvenUmeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten)(Swepub:umu)svbe0001 (författare)
Benz, Roland (författare)
Umeå universitetInstitutionen för molekylärbiologi (Medicinska fakulteten) (creator_code:org_t)

Ingår i:Journal of Biological Chemistry: American Society for Biochemistry and Molecular Biology289:27, s. 18614-186240021-92581083-351X

Av författaren/redakt...: Bárcena-Uribarri ...; Thein, Marcus; Barbot, Mariam; Sans-Serramitjan ...; Bonde, Mari; Mentele, Reinhar ...; visa fler...; Lottspeich, Frie ...; Bergström, Sven; Benz, Roland; visa färre...

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