Structural characterization of the protein cce_0567 from Cyanothece 51142, a metalloprotein associated with nitrogen fixation in the DUF683 family
The genome of many cyanobacacteria contain the sequence for a small protein (<100 amino acids) with a commom "domain of unknown function" grouped into the DUF683 protein family. While the biological function of DUF683 is still not known, their genomic location within nitrogen fixation clusters suggests that DUF683 proteins may play a role in the process. The diurnal cyanobacterium Cyanothece sp. PCC 51142 contains a gene for a protein that fall into the DUF683 family, cce_0567 (78 aa, 9.0 kDa). In an effort to elucidate the biochemical role DUF683 proteins may play in nitrogen fixation, we have determined the first crystal structure for a protein in this family, cce_0567, to 1.84 Å resolution. Cce_0567 crystallized in space group P21 with two protein molecules and one Ni2+ cation per asymmetric unit. The protein is composed of two α-helices from residues P11 to G41 (α1) and L49-E74 (α2) with the second α-helix containing a short 310-helix (Y46 - N48). A four-residue linker (L42 - D45) between the helices allows them to form an anti-parallel bundle that cross over each other towards their termini. In solution it is likely that two molecules of cce_0567 form a rod-like dimer by the stacking interactions of ~1/2 of the protein. Histidine-36 is highly conserved in all known DUF683 proteins and the N2 nitrogen of the H36 side chain of each molecule in the dimer coordinate with Ni2+ in the crystal structure. The divalent cation Ni2+ was titrated into 15N-labelled cce_0567 and chemical shift perturbations were observed only in the 1H-15N HSQC spectra for residues at, or near, the site of Ni2+ binding observed in the crystal structure. There was no evidence for an increase in the size of cce_0567 upon binding Ni2+, even in large molar excess of Ni2+, indicating that a metal was not required for dimer formation. Circular dichroism spectroscopy indicated that cce_0567 was extremely robust, with a melting temperature of ~62ºC that was reversible.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 951815
- Report Number(s):
- PNNL-SA-61462; 14398; TRN: US200913%%137
- Journal Information:
- Biochimica et Biophysica Acta--Proteins and Proteomics, 1794(4):627-633, Vol. 1794, Issue 4
- Country of Publication:
- United States
- Language:
- English
Similar Records
Backbone 1H, 13C, and 15N NMR assignments for the Cyanothece 51142 protein cce_0567: a protein associated with nitrogen fixation in the DUF683 family
Crystal structure of cce_0566 from Cyanothece 51142, a protein associated with nitrogen fixation in the DUF269 family
Related Subjects
AMINO ACIDS
BIOLOGICAL FUNCTIONS
CATIONS
CHEMICAL SHIFT
CRYSTAL STRUCTURE
CYANOBACTERIA
DICHROISM
DIMERS
GENES
MELTING
METALLOPROTEINS
NITROGEN
NITROGEN FIXATION
PROTEINS
RESIDUES
SPACE GROUPS
SPECTRA
SPECTROSCOPY
Metalloprotein
nitrogen fixation
cyanobacteria
circadian rhythms
nickel-binding
Environmental Molecular Sciences Laboratory