NaCl‐activated nucleoside diphosphate kinase from extremely halophilic archaeon, <i>Halobacterium salinarum</i>, maintains native conformation without salt

Ishibashi, Matsujiro; Tokunaga, Hiroko; K, Hiratsuka; Yonezawa, Yasushi; Tsurumaru, Hirohito; Arakawa, Tsutomu; Tokunaga, Masao

doi:10.1016/s0014-5793(01)02292-x

Cited by 47 publications

(66 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, HsNDK from H. salinarum was active and stable in the low-salt conditions, as we have reported previously [6]. Here we demonstrated that HsNDK associates into a hexamer at 3.8 M NaCl and dissociates into a dimer at 0.2 M NaCl reversibly depending on the salt concentrations without converting into enzymatically inactive monomer.…”

Section: Discussionsupporting

confidence: 80%

See 1 more Smart Citation

Secondary and quaternary structural transition of the halophilic archaeon nucleoside diphosphate kinase under high- and low-salt conditions

Ishibashi

2002

FEMS Microbiology Letters

View full text Add to dashboard Cite

Most halophilic enzymes from extremely halophilic archaea are denatured immediately after transfer from high-salt to low-salt medium. However, nucleoside diphosphate kinase (HsNDK) from the extremely halophilic archaeon Halobacterium salinarum seems to be exceptional, since the enzyme exhibited catalytic activity even under the low-salt condition. Here we show the mechanism how HsNDK is active under both high-and low-salt conditions that the HsNDK hexamer in high-salt medium dissociates into a dimer in the low-salt medium without denaturation. The observed change of the subunit structure was accompanied by a large decrease of K-helical content and lowered thermal sensitivity, yet keeping the conformations. This novel hexamer to dimer conversion under high-and low-salt conditions, respectively, seems to be the mechanism by which HsNDK is avoided from the irreversible denaturation.

show abstract

Section: Discussionsupporting

confidence: 80%

“…Since HsNDK is enzymatically active in 0.2 M NaCl [6], we ¢rst examined the secondary structure of HsNDK in both high-and low-salt conditions. The far UV CD spectrum of HsNDK in 3.8 M NaCl showed a secondary structure containing 29% K-helix, 19% L-sheet and 18% L-turns (Fig.…”

Section: Secondary Structures Of Hsndk At High-and Low-salt Concentramentioning

confidence: 99%

Secondary and quaternary structural transition of the halophilic archaeon nucleoside diphosphate kinase under high- and low-salt conditions

Ishibashi

2002

FEMS Microbiology Letters

View full text Add to dashboard Cite

show abstract

“…DnaK itself has MW of 67.4 kDa, from which its alias Hsp 70 was historically coined, but is predicted to migrate as 88.9 kDa in gels. A western blot performed seventeen years ago to study a nucleoside diphosphate kinase [67] also shows a clear ~45 kDa band with the same expression patterns of the DnaK band [68] (Fig. S18).…”

Section: Resultsmentioning

confidence: 99%

Internal RNAs overlapping coding sequences can drive the production of alternative proteins in archaea

et al. 2018

View full text Add to dashboard Cite

Prokaryotic genomes show a high level of information compaction often with different molecules transcribed from the same locus. Although antisense RNAs have been relatively well studied, RNAs in the same strand, internal RNAs (intraRNAs), are still poorly understood. The question of how common is the translation of overlapping reading frames remains open. We address this question in the model archaeon Halobacterium salinarum. In the present work we used differential RNA-seq (dRNA-seq) in H. salinarum NRC-1 to locate intraRNA signals in subsets of internal transcription start sites (iTSS) and establish the open reading frames associated to them (intraORFs). Using C-terminally flagged proteins, we experimentally observed isoforms accurately predicted by intraRNA translation for kef1, acs3 and orc4 genes. We also recovered from the literature and mass spectrometry databases several instances of protein isoforms consistent with intraRNA translation such as the gas vesicle protein gene gvpC1. We found evidence for intraRNAs in horizontally transferred genes such as the chaperone dnaK and the aerobic respiration related cydA in both H. salinarum and Escherichia coli. Also, intraRNA translation evidence in H. salinarum, E. coli and yeast of a universal elongation factor (aEF-2, fusA and eEF-2) suggests that this is an ancient phenomenon present in all domains of life.

show abstract

“…The refolded sample was incubated overnight at 4°C and measured for enzyme activity. Enzymatic activity was assayed as described before [4], except for using a micro-titer plate (the volume of reaction mixture was 0.2 ml) and a plate reader (Benchmark Plus, Bio-Rad). The HsNDK mutant gene of interest thus isolated was re-cloned at NdeI/BamHI site of pET3a to construct pG114R, analyzed by ABI PRISM 3100 Genetic Analyzer (Applied Biosystems) for DNA sequence determination, and used for further characterization.…”

Section: Isolation Expression and Screening Of Hsndk Mutantsmentioning

confidence: 99%

A single Gly114Arg mutation stabilizes the hexameric subunit assembly and changes the substrate specificity of halo‐archaeal nucleoside diphosphate kinase

Ishibashi¹,

Tatsuda²,

Izutsu³

et al. 2007

FEBS Letters

Self Cite

View full text Add to dashboard Cite

Nucleoside diphosphate kinase from extremely halophilic archaeon (HsNDK) requires above 2 M NaCl concentration for in vitro refolding. Here an attempt was made to isolate mutations that allow HsNDK to refold in low salt media. Such a screening resulted in isolation of an HsNDK mutant, Gly114Arg, which efficiently refolded in the presence of 1 M NaCl. This mutant, unlike the wild type enzyme, was expressed in Escherichia coli as an active form. The residue 114 is in close proximity to Glu155 of the neighboring subunit in the three dimensional hexameric structure of the HsNDK. It is thus possible that the attractive electrostatic interactions occur between Arg114 and Glu155 in the mutant HsNDK, stabilizing the hexameric subunit assembly.

show abstract

NaCl‐activated nucleoside diphosphate kinase from extremely halophilic archaeon, Halobacterium salinarum, maintains native conformation without salt

Cited by 47 publications

References 22 publications

Secondary and quaternary structural transition of the halophilic archaeon nucleoside diphosphate kinase under high- and low-salt conditions

Secondary and quaternary structural transition of the halophilic archaeon nucleoside diphosphate kinase under high- and low-salt conditions

Internal RNAs overlapping coding sequences can drive the production of alternative proteins in archaea

A single Gly114Arg mutation stabilizes the hexameric subunit assembly and changes the substrate specificity of halo‐archaeal nucleoside diphosphate kinase

Contact Info

Product

Resources

About