The crystal structure of auracyanin A at 1.85 Å resolution: the structures and functions of auracyanins A and B, two almost identical “blue” copper proteins, in the photosynthetic bacterium Chloroflexus aurantiacus (original) (raw)

Abstract

Auracyanins A and B are two closely similar “blue” copper proteins produced by the filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus. Both proteins have a water-soluble 140-residue globular domain, which is preceded in the sequence by an N-terminal tail. The globular domains of auracyanins A and B have sequences that are 38% identical. The sequences of the N-terminal tails, on the other hand, are distinctly different, suggesting that auracyanins A and B occupy different membrane sites and have different functions. The crystal structure of auracyanin A has been solved and refined at 1.85 Å resolution. The polypeptide fold is similar to that of auracyanin B (Bond et al. in J Mol Biol 306:47–67, 2001), but the distribution of charged and polar residues on the molecular surface is different. The Cu-site dimensions of the two auracyanins are identical. This is unexpected, since auracyanin A has a shorter polypeptide loop between two of the Cu-binding residues, and the two proteins have significantly different EPR, UV–visible and resonance Raman spectra. The genes for the globular domains of auracyanins A and B have been cloned in a bacterial expression system, enabling purification of large quantities of protein. It is shown that auracyanin A is expressed only when C. aurantiacus cells are grown in light, whereas auracyanin B is expressed under dark as well as light conditions. The inference is that auracyanin A has a function in photosynthesis, and that auracyanin B has a function in aerobic respiration.

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Abbreviations

BCIP:

5-Bromo-4-chloro-3-indolyl phosphate

Chx:

Chloroflexus aurantiacus

DFT:

Density functional theory

DPI:

Diffraction data precision indicator

ESU:

Estimated standard uncertainty

EXAFS:

Extended X-ray absorption fine structure

MALDI-TOF:

Matrix-assisted laser desorption ionization-time-of-flight

MW:

Molecular weight

NBT:

Nitro blue tetrazolium chloride

RMS:

Root-mean-square

XANES:

X-ray absorption near-edge structure

XAS:

X-ray absorption spectroscopy

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Acknowledgments

Support from the Australian Research Council (Grant DP0208320 to JMG and HCF), the Molecular Biochemistry program of the National Science Foundation (Grant MCB0646621 to REB), and the Australian Access to Major Research Facilities program (travel grant to HHH and HCF) is gratefully acknowledged. The Stanford Synchrotron Research Laboratory is a US national user facility operated by Stanford University on behalf of the US Department of Energy, Office of Basic Energy Sciences. We thank Stephen Graham for recording X-ray diffraction data, and Christopher Dennison, Graham George and Edward Solomon for helpful discussions.

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Authors and Affiliations

1. Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ, 85287, USA
   Melissa C. del Rosario
2. School of Chemistry, University of Sydney, Sydney, NSW, 2006, Australia
   Hugh H. Harris
3. Departments of Biology and Chemistry, Washington University in St Louis, St Louis, MO, 63130, USA
   Robert E. Blankenship
4. School of Molecular and Microbial Biosciences, University of Sydney, Sydney, NSW, 2006, Australia
   Mihwa Lee, J. Mitchell Guss & Hans C. Freeman

Authors

1. Mihwa Lee
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2. Melissa C. del Rosario
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3. Hugh H. Harris
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4. Robert E. Blankenship
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5. J. Mitchell Guss
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6. Hans C. Freeman
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Corresponding authors

Correspondence toRobert E. Blankenship or J. Mitchell Guss.

Additional information

H. C. Freeman passed away in November 2008 shortly before he could submit the final manuscript. The remaining authors dedicate this paper to his memory in recognition of his pioneering research on Cu proteins.

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Lee, M., del Rosario, M.C., Harris, H.H. et al. The crystal structure of auracyanin A at 1.85 Å resolution: the structures and functions of auracyanins A and B, two almost identical “blue” copper proteins, in the photosynthetic bacterium Chloroflexus aurantiacus .J Biol Inorg Chem 14, 329–345 (2009). https://doi.org/10.1007/s00775-009-0473-0

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• Received: 27 November 2008
• Accepted: 18 January 2009
• Published: 04 February 2009
• Issue Date: March 2009
• DOI: https://doi.org/10.1007/s00775-009-0473-0

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