Part II: Diffraction from two-dimensional cholera toxin crystals bound to their receptors in a lipid monolayer

C. E. Miller; Jarek P. Majewski; E. B. Watkins; M. Weygand; T. L. Kuhl

doi:10.1529/biophysj.107.120808

Part II: Diffraction from two-dimensional cholera toxin crystals bound to their receptors in a lipid monolayer

C. E. Miller
, Jarek P. Majewski
, E. B. Watkins
, M. Weygand
, T. L. Kuhl

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

The structure of cholera toxin (CTAB₅) bound to its putative ganglioside receptor, galactosyl-N-acetylgalactosaminyl (N-acetyl-neuraminyl) galactosylglucosylceramide (GM₁), in a lipid monolayer at the air-water interface has been studied utilizing grazing incidence x-ray diffraction. Cholera toxin is one of very few proteins to be crystallized in two dimensions and characterized in a fully hydrated state. The observed grazing incidence x-ray diffraction Bragg peaks indicated cholera toxin was ordered in a hexagonal lattice and the order extended 600-800 Å. The pentameric binding portion of cholera toxin (CTB₅) improved in-plane ordering over the full toxin (CTAB₅) especially at low pH. Disulfide bond reduction (activation of the full toxin) also increased the protein layer ordering. These findings are consistent with A-subunit flexibility and motion, which cause packing inefficiencies and greater disorder of the protein layer. Corroborative out-of-plane diffraction (Bragg rod) analysis indicated that the scattering units in the cholera layer with CTAB₅ shortened after disulfide bond reduction of the A subunit. These studies, together with Part I results, revealed key changes in the structure of the cholera toxin-lipid system under different pH conditions.

Original language	English
Pages (from-to)	641-647
Number of pages	7
Journal	Biophysical Journal
Volume	95
Issue number	2
DOIs	https://doi.org/10.1529/biophysj.107.120808
State	Published - Jul 15 2008
Externally published	Yes

Funding

The Los Alamos Neutron Science Center at the Los Alamos National Laboratory (LANL) is funded by the U.S. Department of Energy (DOE) under Contract W-7405-ENG-36. J.M and C.E.M. thank the LANL-Laboratory Directed Research and Development program, DOE Office of Science (Basic Energy Sciences) for financial support. C.E.M. acknowledges support from the LANL Director's Post-Doctoral Fellowship and the Institute for Complex Adaptive Matter. T.L.K. thanks the Jeff and Dianne Child/Steve Whitaker Fund for Distinguished Teaching and Scholarship for financial support.

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title = "Part II: Diffraction from two-dimensional cholera toxin crystals bound to their receptors in a lipid monolayer",

abstract = "The structure of cholera toxin (CTAB5) bound to its putative ganglioside receptor, galactosyl-N-acetylgalactosaminyl (N-acetyl-neuraminyl) galactosylglucosylceramide (GM1), in a lipid monolayer at the air-water interface has been studied utilizing grazing incidence x-ray diffraction. Cholera toxin is one of very few proteins to be crystallized in two dimensions and characterized in a fully hydrated state. The observed grazing incidence x-ray diffraction Bragg peaks indicated cholera toxin was ordered in a hexagonal lattice and the order extended 600-800 {\AA}. The pentameric binding portion of cholera toxin (CTB5) improved in-plane ordering over the full toxin (CTAB5) especially at low pH. Disulfide bond reduction (activation of the full toxin) also increased the protein layer ordering. These findings are consistent with A-subunit flexibility and motion, which cause packing inefficiencies and greater disorder of the protein layer. Corroborative out-of-plane diffraction (Bragg rod) analysis indicated that the scattering units in the cholera layer with CTAB5 shortened after disulfide bond reduction of the A subunit. These studies, together with Part I results, revealed key changes in the structure of the cholera toxin-lipid system under different pH conditions.",

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AU - Weygand, M.

AU - Kuhl, T. L.

PY - 2008/7/15

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AB - The structure of cholera toxin (CTAB5) bound to its putative ganglioside receptor, galactosyl-N-acetylgalactosaminyl (N-acetyl-neuraminyl) galactosylglucosylceramide (GM1), in a lipid monolayer at the air-water interface has been studied utilizing grazing incidence x-ray diffraction. Cholera toxin is one of very few proteins to be crystallized in two dimensions and characterized in a fully hydrated state. The observed grazing incidence x-ray diffraction Bragg peaks indicated cholera toxin was ordered in a hexagonal lattice and the order extended 600-800 Å. The pentameric binding portion of cholera toxin (CTB5) improved in-plane ordering over the full toxin (CTAB5) especially at low pH. Disulfide bond reduction (activation of the full toxin) also increased the protein layer ordering. These findings are consistent with A-subunit flexibility and motion, which cause packing inefficiencies and greater disorder of the protein layer. Corroborative out-of-plane diffraction (Bragg rod) analysis indicated that the scattering units in the cholera layer with CTAB5 shortened after disulfide bond reduction of the A subunit. These studies, together with Part I results, revealed key changes in the structure of the cholera toxin-lipid system under different pH conditions.

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Part II: Diffraction from two-dimensional cholera toxin crystals bound to their receptors in a lipid monolayer

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