• 한국자기공명학회논문지
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• 제19권3호
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• pp.106-111
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• 2015

Helicobacter pylori (H. pylori) survives in acidic and fluctuating pH conditions of the stomach. The pH effect on H. pylori proteins is important for the advanced understanding of its evolution and viability, although this bacterium has the molecular machinery that neutralizes the acidic condition. HP1492 is known as a conserved NifU-like protein from H. pylori. NifU is a nitrogen fixation protein that mediates the transfer of iron-sulfur (Fe-S) cluster to iron-sulfur proteins like ferredoxin. Commonly, the monomeric reduced state of NifU can be converted to the dimeric oxidized state by intermolecular disulfide bond formation. Because it remains unclear that HP1492 actually behaves as known NifU protein, we first found that this protein can adopt both oxidized and reduced forms using size exclusion chromatography. Circular dichroism experiment showed that HP1492 is relatively well-structured at pH 6.5, compared to other pH conditions. On the basis of the backbone resonance assignment of HP1492, we further characterized the residues that are sensitive to pH using NMR spectroscopy. These residues showing large chemical shift changes could be mapped onto the secondary structure of the protein. Our results could provide the foundation for structural and biophysical studies on a wide spectrum of NifU proteins.

• Genomics & Informatics
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• 제8권4호
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• pp.170-176
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• 2010

The Ribonucleotide reductases (RNR) are essential enzymes that catalyze the conversion of nucleotides to deoxynucleotides in DNA replication and repair in all living organisms. The RNRs operate by a free radical mechanism but differ in the composition of subunit, cofactor required and regulation by allostery. Based on these differences the RNRs are classified into three classesclass I, class II and class III which depend on oxygen, adenosylcobalamin and S-adenosylmethionine with an iron sulfur cluster respectively for radical generation. In this article thirty seven sequences belonging to each of the three classes of RNR were analyzed by using various tools of bioinformatics. Phylogenetic analysis, dot-plot comparisons and motif analysis was done to identify a number of differences in the three classes of RNRs. In this research article, we have attempted to decipher evolutionary relationship between the three classes of RNR by using bioinformatics approach.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.134-139
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• 2004

Hybrid cluster protein (HCP) was investigated because of its unique iron-sulfur clusters, which have been found in bacteria and archaea. Here, HCP homologous proteins from the third domain, 'eukarya'(3 amitochondriate protozoans, Giardia lamblia, Entamoeba histolytica, and Trichomonas vaginalis), were identified. All three amitochondriate protozoan HCPs (GlHCP, EhHCP, and TvHCP) belonged to Class I on the basis of two key characters, the cysteine spacing, Cys-(Xaa)₂Cys(Xaa)/sub 7-8/-Cys(Xaa)/sub 5/-Cys, and the absence of N-terminal deletion characteristic to the Class III. In phylogenetic analysis performed with amino acid sequences of 3 eukaryal, 5 bacterial, and 4 archaeal HCPs, the maximum likelihood (ML) tree indicated that TvHCP was clustered with Class I HCPs, whereas the other two HCPs (GlHCP and EhHCP) formed an independent clade with a high bootstrapping value (96%) not belonging to any previously recognized HCP class. In spite of the relatively lower bootstrapping value (61%), the position of the new eukaryal GlHCP-EhHCP clade was close to Class I, including the TvHCP, and Classes II and III were closely related with each other. The finding of eukaryal HCPs would help to understand the evolutionary history of HCP.