Tobacco small heat-shock protein, NtHSP18.2, has broad substrate range as a molecular chaperone

Kim, Keun Pill; Joe, Myung Kook; Hong, Choo Bong

doi:10.1016/j.plantsci.2004.05.043

Cited by 9 publications

(12 citation statements)

References 22 publications

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“…Its members share a conserved e~-crystallin domain in the carboxy terminal, and show diversity in the amino terminal (Caspers et al, 1995). Studies of sHsps from various organisms have shown that they bind to their substrates stably and protect them, in an A-P-independent manner, from thermal or chemical aggregation (Ehrnsperger et al, 1997;Wang and gpector, 2000;Abdulle et al, 2002;Stromer et al 2003;Basha et al, 2004b;Fu and Chang, 2004;Kim et al, 2004). This chaperone activity is likely related to the stress tolerance of cells (Joe et al, 2000;Park and Hong, 2002).…”

Section: Discussionmentioning

confidence: 99%

“…Previously, we analyzed the functional modes of NtHSP18.2 and NtHSP18.3 (Kim et al, 2004;Yu, 2004). Here, we describe the in vitro chaperone activity of NtHSP1 7.6, another tobacco sHsp.…”

Section: Discussionmentioning

confidence: 99%

“…After heating to 95~ for 3 min, PCR was performed in a thermal cycler (Perkin-EImer, USA) for 30 cycles (95~ 30 s; 50~ 30 s; and 72~ 1 min). The amplified product was digested with Sacl and Sphl, and ligated into the pBAD expression vector (Kim et al, 2004) at the 5acl and Sphl cloning sites. The sequence of the cloned coding region was confirmed by dideoxy chain termination sequencing (Sanger et al, 1977), using Sequenase version 2.0 (United States Biochemical, USA).…”

Section: Genomic Clone Nthsp176mentioning

confidence: 99%

“…Early studies revealed that, among those clones, NtHSP18.2 and NtHSP18.3 protected a broad range of proteins from heat shock (Kim et al, 2004;Yu, 2004). In the present report, we show that another sHsp from tobacco, NtHSP17.6, also has in vitro chaperone activity.…”

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Functional mode of NtHSP17.6, a cytosolic small heat-shock protein fromNicotiana tabacum

et al. 2005

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Small heat-shock proteins (sHsps) are ubiquitous stress proteins with molecular chaperone activity. They share characteristic homology with the @-crystallin protein of the mammalian eye lens as well as being ATP-independent in their chaperone activity. We isolated a clone for a cytosolic class ! sHsp, NtHSP17.6, from Nicotiana tabacum, and analyzed its functional mode for such activity. Following its transformation into Escherichia coli and its over-expression, NtHSP17.6 was purified and examined in vitro. This purified NtHSP17.6 exhibited typical chaperone activity in a lightscattering test. It was enable to protect a model substrate, firefly luciferase, from heat-induced aggregation. Nondenaturing PAGE showed that NtHSP17.6 formed a dodecamer in its native conformation, and was bound to its substrate under heat stress. A labeling test with bis-ANS indicated that this binding might be linked to newly exposed hydrophobic sites of the NtHSP17.6 complexes during heat shock. Based on these data, we suggest that NtHSP17.6 is a molecular chaperone that functions as a dodecamer in a heat-induced manner.

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Section: Discussionmentioning

confidence: 99%

“…Previously, we analyzed the functional modes of NtHSP18.2 and NtHSP18.3 (Kim et al, 2004;Yu, 2004). Here, we describe the in vitro chaperone activity of NtHSP1 7.6, another tobacco sHsp.…”

Section: Discussionmentioning

confidence: 99%

Section: Genomic Clone Nthsp176mentioning

confidence: 99%

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Functional mode of NtHSP17.6, a cytosolic small heat-shock protein fromNicotiana tabacum

et al. 2005

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“…Luc and CS are heat-labile proteins that have been commonly used as model substrates for in vitro molecular chaperone assays under high temperature (6,13). Luc and CS both showed a rapid increase in light scattering at 340 nm upon heat treatment at 42 o C for Luc and 45 o C for CS.…”

Section: Resultsmentioning

confidence: 99%

Tobacco mitochondrial small heat shock protein NtHSP24.6 adopts a dimeric configuration and has a broad range of substrates

Kim

Yu²,

Park³

et al. 2011

BMB Reports

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There is a broad range of different small heat shock proteins (sHSPs) that have diverse structural and functional characteristics. To better understand the functional role of mitochondrial sHSP, NtHSP24.6 was expressed in Escherichia coli with a hexahistidine tag and purified. The protein was analyzed by non-denaturing PAGE, chemical cross-linking and size exclusion chromatography and the H6NtHSP24.6 protein was found to form a dimer in solution. The in vitro functional analysis of H6NtHSP24.6 using firefly luciferase and citrate synthase demonstrated that this protein displays typical molecular chaperone activity. When cell lysates of E. coli were heated after the addition of H6NtHSP24.6, a broad range of proteins from 10 to 160 kD in size remained in the soluble state. These results suggest that NtHSP24.6 forms a dimer and can function as a molecular chaperone to protect a diverse range of proteins from thermal aggregation. [BMB reports 2011; 44(12): 816-820]

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Enhanced tolerance against freezing stress inEscherichia coli cells expressing an algal cyclophilin gene

Cho

2007

Biotechnol. Bioprocess Eng.

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Tobacco small heat-shock protein, NtHSP18.2, has broad substrate range as a molecular chaperone

Cited by 9 publications

References 22 publications

Functional mode of NtHSP17.6, a cytosolic small heat-shock protein fromNicotiana tabacum

Functional mode of NtHSP17.6, a cytosolic small heat-shock protein fromNicotiana tabacum

Tobacco mitochondrial small heat shock protein NtHSP24.6 adopts a dimeric configuration and has a broad range of substrates

Enhanced tolerance against freezing stress inEscherichia coli cells expressing an algal cyclophilin gene

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