In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 Å Resolution

V. Cherezov, E. Yamashita, Wei Liu, M. Zhalnina, W. A. Cramer, M. Caffrey

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Crystals of the apo form of the vitamin B12 and colicin receptor, BtuB, that diffract to 1.95 Å have been grown by the membrane-based in meso technique. The structure of the protein differs in several details from that of its counterpart grown by the more traditional, detergent-based (in surfo) method. Some of these differences include (i) the five N-terminal residues are resolved in meso, (ii) residues 57-62 in the hatch domain and residues 574-581 in loop 21-22 are disordered in meso and are ordered in surfo, (iii) residues 278-287 in loop 7-8 are resolved in meso, (iv) residues 324-331 in loop 9-10, 396-411 in loop 13-14, 442-458 in loop 15-16 and 526-541 in loop 19-20 have large differences in position between the two crystal forms, as have residues 86-96 in the hatch domain, and (v) the conformation of residues 6 and 7 in the Ton box (considered critical to signal transduction and substrate transport) are entirely different in the two structures. Importantly, the in meso orientation of residues 6 and 7 is similar to that of the vitamin B12-charged state. These data suggest that the "substrate-induced" 180° -rotation of residues 6 and 7 reported in the literature may not be a unique signalling event. The extent to which these findings agree with structural, dynamic and functional insights gleaned from site-directed spin labelling and electron paramagnetic resonance measurements is evaluated. Packing in in meso grown crystals is dense and layered, consistent with the current model for crystallogenesis of membrane proteins in lipidic mesophases. Layered packing has been used to locate the transmembrane hydrophobic surface of the protein. Generally, this is consistent with tryptophan, tyrosine, lipid and Cα B-factor distributions in the protein, and with predictions based on transfer free energy calculations.

Original languageEnglish (US)
Pages (from-to)716-734
Number of pages19
JournalJournal of Molecular Biology
Volume364
Issue number4
DOIs
StatePublished - Dec 8 2006
Externally publishedYes

Fingerprint

Vitamin B 12
Membrane Proteins
Colicins
Energy Transfer
Electron Spin Resonance Spectroscopy
Tryptophan
Detergents
Tyrosine
Signal Transduction
Proteins
Lipids
Membranes
3'-(1-butylphosphoryl)adenosine

Keywords

  • colicin
  • cubic phase
  • membrane protein crystals
  • vitamin B
  • X-ray diffraction

ASJC Scopus subject areas

  • Virology

Cite this

In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 Å Resolution. / Cherezov, V.; Yamashita, E.; Liu, Wei; Zhalnina, M.; Cramer, W. A.; Caffrey, M.

In: Journal of Molecular Biology, Vol. 364, No. 4, 08.12.2006, p. 716-734.

Research output: Contribution to journalArticle

Cherezov, V, Yamashita, E, Liu, W, Zhalnina, M, Cramer, WA & Caffrey, M 2006, 'In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 Å Resolution', Journal of Molecular Biology, vol. 364, no. 4, pp. 716-734. https://doi.org/10.1016/j.jmb.2006.09.022
Cherezov, V. ; Yamashita, E. ; Liu, Wei ; Zhalnina, M. ; Cramer, W. A. ; Caffrey, M. / In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 Å Resolution. In: Journal of Molecular Biology. 2006 ; Vol. 364, No. 4. pp. 716-734.
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T1 - In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 Å Resolution

AU - Cherezov, V.

AU - Yamashita, E.

AU - Liu, Wei

AU - Zhalnina, M.

AU - Cramer, W. A.

AU - Caffrey, M.

PY - 2006/12/8

Y1 - 2006/12/8

N2 - Crystals of the apo form of the vitamin B12 and colicin receptor, BtuB, that diffract to 1.95 Å have been grown by the membrane-based in meso technique. The structure of the protein differs in several details from that of its counterpart grown by the more traditional, detergent-based (in surfo) method. Some of these differences include (i) the five N-terminal residues are resolved in meso, (ii) residues 57-62 in the hatch domain and residues 574-581 in loop 21-22 are disordered in meso and are ordered in surfo, (iii) residues 278-287 in loop 7-8 are resolved in meso, (iv) residues 324-331 in loop 9-10, 396-411 in loop 13-14, 442-458 in loop 15-16 and 526-541 in loop 19-20 have large differences in position between the two crystal forms, as have residues 86-96 in the hatch domain, and (v) the conformation of residues 6 and 7 in the Ton box (considered critical to signal transduction and substrate transport) are entirely different in the two structures. Importantly, the in meso orientation of residues 6 and 7 is similar to that of the vitamin B12-charged state. These data suggest that the "substrate-induced" 180° -rotation of residues 6 and 7 reported in the literature may not be a unique signalling event. The extent to which these findings agree with structural, dynamic and functional insights gleaned from site-directed spin labelling and electron paramagnetic resonance measurements is evaluated. Packing in in meso grown crystals is dense and layered, consistent with the current model for crystallogenesis of membrane proteins in lipidic mesophases. Layered packing has been used to locate the transmembrane hydrophobic surface of the protein. Generally, this is consistent with tryptophan, tyrosine, lipid and Cα B-factor distributions in the protein, and with predictions based on transfer free energy calculations.

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KW - colicin

KW - cubic phase

KW - membrane protein crystals

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KW - X-ray diffraction

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