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Use of nonelectroly...
Use of nonelectrolytes reveals the channel size and oligomeric constitution of the Borrelia burgdorferi P66 porin
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- Bárcena-Uribarri, Iván (författare)
- Universität Würzburg, Germany
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- Thein, Marcus (författare)
- Universität Würzburg and Jacobs University Bremen, Germany
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- Maier, Elke (författare)
- Universität Würzburg, Germany
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- Bonde, Mari (författare)
- Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten)
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- Bunikis, Ignas, 1981- (författare)
- Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten)
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- Bergström, Sven, 1950- (författare)
- Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten)
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- Benz, Roland (författare)
- Universität Würzburg, Germany
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(creator_code:org_t)
- Engelska.
- Relaterad länk:
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https://umu.diva-por... (primary) (Raw object)
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- The outer membrane protein P66 of the Lyme disease spirochete Borrelia burgdorferi is capable of pore formation with an atypical high single-channel conductance of 11 nS in 1 M KCl. We studied in a non-theoretical manner the diameter of the P66 channel by analyzing its single-channel conductance in black lipid bilayers in the presence of different nonelectrolytes with known hydrodynamic radii. Furthermore, we calculated the filling of the channel with these nonelectrolytes and the results revealed that nonelectrolytes with hydrodynamic radii of 0.34 nm or smaller pass through the pore, whereas neutral molecules with greater radii only partially filled the channel or were not able to enter it at all. Thus, the diameter of the P66 entrance was determined to be ≤ 1.9 nm with a constriction site diameter of about 0.7 nm. Furthermore, the P66-induced membrane conductance could be blocked by 80-90% after addition of the nonelectrolytes PEG 400, PEG 600 and maltohexaose in the low millimolar range. Interestingly, the analysis of the power density spectra of P66 after blockage with nonelectrolytes revealed no chemical interaction responsible for channel block. The blockage of one P66 single-channel conductance unit of 11 nS occurred by seven subconducting states, thus indicating a heptameric organization of the P66 oligomer. This organization of P66 as a heptamer was confirmed by Blue Native PAGE and immunoblot analysis, which demonstrated that P66 forms a complex with a mass of approximately 460 kDa.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Nyckelord
- nutrient uptake
- outer membrane transport
- Lyme disease
- porin radius determination
- black lipid bilayer
- Molecular biology
- Molekylärbiologi
- molekylärbiologi
- Molecular Biology
Publikations- och innehållstyp
- vet (ämneskategori)
- ovr (ämneskategori)