scholarly journals The Archaeal Small Heat Shock Protein Hsp17.6 Protects Proteins from Oxidative Inactivation

2021 ◽  
Vol 22 (5) ◽  
pp. 2591
Author(s):  
Pengfei Ma ◽  
Jie Li ◽  
Lei Qi ◽  
Xiuzhu Dong
Keyword(s):  
Heat Shock ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Small Heat Shock Protein ◽  
Phase Cell ◽  
Molecular Weights ◽  
Oxidative Inactivation ◽  
Methionine Residues ◽  
Shock Proteins ◽  
And Function

Small heat shock proteins (sHsps) are widely distributed among various types of organisms and function in preventing the irreversible aggregation of thermal denaturing proteins. Here, we report that Hsp17.6 from Methanolobus psychrophilus exhibited protection of proteins from oxidation inactivation. The overexpression of Hsp17.6 in Escherichia coli markedly increased the stationary phase cell density and survivability in HClO and H2O2. Treatments with 0.2 mM HClO or 10 mM H2O2 reduced malate dehydrogenase (MDH) activity to 57% and 77%, whereas the addition of Hsp17.6 recovered the activity to 70–90% and 86–100%, respectively. A similar effect for superoxide dismutase oxidation was determined for Hsp17.6. Non-reducing sodium dodecyl sulfate polyacrylamide gel electrophoresis assays determined that the Hsp17.6 addition decreased H2O2-caused disulfide-linking protein contents and HClO-induced degradation of MDH; meanwhile, Hsp17.6 protein appeared to be oxidized with increased molecular weights. Mass spectrometry identified oxygen atoms introduced into the larger Hsp17.6 molecules, mainly at the aspartate and methionine residues. Substitution of some aspartate residues reduced Hsp17.6 in alleviating H2O2- and HClO-caused MDH inactivation and in enhancing the E. coli survivability in H2O2 and HClO, suggesting that the archaeal Hsp17.6 oxidation protection might depend on an “oxidant sink” effect, i.e., to consume the oxidants in environments via aspartate oxidation

scholarly journals Heat Shock Protein-Mediated Resistance to High Hydrostatic Pressure in Escherichia coli

2004 ◽  
Vol 70 (5) ◽  
pp. 2660-2666 ◽  
Author(s):  
Abram Aertsen ◽  
Kristof Vanoirbeek ◽  
Philipp De Spiegeleer ◽  
Jan Sermon ◽  
Kristel Hauben ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
High Hydrostatic Pressure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
E Coli ◽  
Shock Proteins

ABSTRACT A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon (dnaK, lon, clpPX), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i) the expression of rpoH, encoding the heat shock-specific sigma factor σ32, was also induced by high pressure; (ii) heat shock rendered E. coli significantly more resistant to subsequent high-pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii) basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels.

Effect of heat shock on protein synthesis in flax rust uredosporelings

1985 ◽  
Vol 63 (11) ◽  
pp. 2069-2076 ◽  
Author(s):  
Maichael Shaw ◽  
Rosalinda Boasson ◽  
Leroy Scrubb
Keyword(s):  
Heat Shock ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Heat Stroke ◽  
Sodium Dodecyl ◽  
Axenic Culture ◽  
One Dimensional ◽  
Melampsora Lini ◽  
Molecular Masses ◽  
Shock Proteins

In uredosporelings of Melampsora lini (Ehrenb.) Lév. germinated for 3 h, uptake of [35S]methionine and its incorporation into protein were depressed during a 2-h heat shock induced by transfer from 17 ± 0.5 to 31 °C. Spectrophotometric scans of fluorograms, prepared after one-dimensional sodium dodecyl sulphate – polyacrylamide gel electrophoresis of aliquots of protein extracts containing equal numbers of disintegrations per minute (1 dps = 1 Bq) in protein, showed that heat shock induced statistically significant changes in the relative degrees of incorporation of [35S]methionine into 15 polypeptide bands. Increased labelling occurred in seven bands, which appear to be heat shock proteins with apparent molecular masses of 84, 71, 43.5, 30.5, 19.5, 18, and 17 kD. Decreased labelling occurred in eight bands, which appear to be heat stroke proteins with apparent molecular masses of 56, 54, 48, 46, 34, 32, 31.5, and 14 kD. When [35S]methionine was administered during heat shock at 31 °C the same pattern of polypeptide labelling was observed in extracts made immediately without return to 17 °C and in extracts made after uredosporelings were returned to 17 °C for 24 h. Thus label incorporated into each polypeptide during heat shock was retained for at least 24 h after the return to 17 °C. Administration of [35S]methionine at 17 °C after completion of the heat shock showed that the "normal" or "nonshock" pattern of labelling began to be resumed within 2 h after transfer from 31 to 17 °C. The results are discussed in relation to the effect of heat shock in promoting the initiation and growth of mycelial colonies from uredosporelings in axenic culture.

scholarly journals Identification and Expression Analysis of Four Small Heat Shock Protein Genes in Cigarette Beetle, Lasioderma serricorne (Fabricius)

Insects ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 139 ◽  
Author(s):  
Wen-Jia Yang ◽  
Kang-Kang Xu ◽  
Yu Cao ◽  
Yong-Lu Meng ◽  
Yan Liu ◽  
...  
Keyword(s):  
Heat Shock ◽  
Fat Body ◽  
Small Heat Shock Protein ◽  
Transcript Level ◽  
Lasioderma Serricorne ◽  
Transcript Levels ◽  
Cigarette Beetle ◽  
Molecular Weights ◽  
Shock Proteins ◽  
High Expression Level

Small heat shock proteins (sHsps) are molecular chaperones that play crucial roles in the stress adaption of insects. In this study, we identified and characterized four sHsp genes (LsHsp19.4, 20.2, 20.3, and 22.2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The four cDNAs encoded proteins of 169, 180, 181, and 194 amino acids with molecular weights of 19.4, 20.2, 20.3, and 22.2 kDa, respectively. The four LsHsp sequences possessed a typical sHsp domain structure. Quantitative real-time PCR analyses revealed that LsHsp19.4 and 20.3 transcripts were most abundant in pupae, whereas the transcript levels of LsHsp20.2 and 22.2 were highest in adults. Transcripts of three LsHsp genes were highly expressed in the larval fat body, whereas LsHsp20.2 displayed an extremely high expression level in the gut. Expression of the four LsHsp genes was dramatically upregulated in larvae exposed to 20-hydroxyecdysone. The majority of the LsHsp genes were significantly upregulated in response to heat and cold treatments, while LsHsp19.4 was insensitive to cold stress. The four genes were upregulated when challenged by immune triggers (peptidoglycan isolated from Staphylococcus aureus and from Escherichia coli 0111:B4). Exposure to CO2 increased LsHsp20.2 and 20.3 transcript levels, but the LsHsp19.4 transcript level declined. The results suggest that different LsHsp genes play important and distinct regulatory roles in L. serricorne development and in response to diverse stresses.

Effect of elevated temperatures on heat shock protein and ribulose-1,5-bisphosphate carboxylase gene expression in Brassica napus

1990 ◽  
Vol 68 (3) ◽  
pp. 609-615 ◽  
Author(s):  
J. R. Halle ◽  
S. Ghosh ◽  
E. B. Dumbroff ◽  
J. J. Heikkila
Keyword(s):  
Gene Expression ◽  
Brassica Napus ◽  
Heat Shock ◽  
Elevated Temperatures ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Immunoblot Analysis ◽  
Bisphosphate Carboxylase ◽  
Molecular Masses ◽  
Shock Proteins

Leaf segments of Brassica napus were exposed to 22, 35, 38, or 40 °C for up to 4 h. Analysis of radiolabeled proteins by two-dimensional sodium dodecyl sulfate – polyacrylamide gel electrophoresis and fluorography revealed two major groups of heat shock proteins (HSPs). One group comprised HSPs 70, 76, and 87, with (pIs) isoelectric points ranging from 5.7 to 6.1, whereas the second group had molecular masses ranging from 23 to 16 kilodaltons (kDa) and pIs from 5.6 to 6.9. Immunoblot analysis using antibodies directed against the large (RLSU) and small (RSSU) subunits of ribulose-1,5-bisphosphate carboxylase (RUBISCO) showed that increasing temperatures from 35 to 38° or 40 °C for the duration of thermal stress (i.e., from 1 to 5 h) did not affect levels of the RSSU (15 kDa), whereas levels of the RLSU (52 kDa) fell sharply. Nevertheless, the activity of RUBISCO was not adversely affected at 38 °C for periods of up to 5 h. Northern blot analysis revealed that the increase observed in HSP 70 synthesis during heat shock may be transcriptionally regulated, but the decrease in the RLSU was not accompanied by a corresponding reduction in levels of its mRNA.Key words: Brassica, heat shock, ribulose-1,5-bisphosphate carboxylase, gene expression.

Low Molecular Weight Trypsin-Plasmin Inhibitors Isolated from Papain Treated Urinary Trypsin Inhibitor

1982 ◽  
Vol 47 (01) ◽  
pp. 014-018 ◽  
Author(s):  
H Sumi ◽  
N Toki ◽  
S Takasugi ◽  
S Maehara ◽  
M Maruyama ◽  
...  
Keyword(s):  
Trypsin Inhibitor ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Chromatography ◽  
Urinary Trypsin Inhibitor ◽  
Plasmin Activity ◽  
Molecular Weights ◽  
Plasmin Inhibitors

SummaryPapain treatment of human urinary trypsin inhibitor (UTI67; mol. wt. 43,000 by SDS-polyacrylamide gel electrophoresis, specific activity 1,897 U/mg protein) produced four new protease inhibitors, which were highly purified by gel chromatography on Sephadex G-100 and isoelectric focusing. The purified inhibitors (UTI26, UTI9-I, UTI9-II, and UTI9-III) were shown to be homogeneous by polyacrylamide disc gel electrophoresis, and had apparent molecular weights of 26,000, 9,000, 9,000, and 9,800, respectively, by sodium dodecyl sulfate gel electrophoresis. During enzymatic degradation of UTI67, the amino acid compositions changed to more basic, and the isoelectric point increased from pH 2.0 (UTI67) to pHs 4.4, 5.2, 6.6, and 8.3 (UTI26, UTI9-I, UTI9-II, and UTI9-III), respectively. Both the parent and degraded inhibitors had anti-plasmin activity as well as antitrypsin and anti-chymotrypsin activities. Much higher anti-plasmin/anti-trypsin and anti-plasmin/anti-chymotrypsin activities were observed in the degraded inhibitors than in the parent UTI67. They competitively inhibited human plasmin with Ki values of 1.13 X 10-7 - 2.12 X 10-6 M (H-D-Val-Leu-Lys-pNA substrate). The reactions were very fast and the active site of the inhibitors to plasmin was thought to be different from that to trypsin or chymotrypsin.

Effect of detergents and chemicals on purified vaccinia virus: analysis by SDS polyacrylamide gel electrophoresis and electron microscopy

1977 ◽  
Vol 23 (3) ◽  
pp. 240-252 ◽  
Author(s):  
J. Boisvert ◽  
T. Yamamoto
Keyword(s):  
Electron Microscopy ◽  
Vaccinia Virus ◽  
Gel Electrophoresis ◽  
Alkali Treatment ◽  
Guanidine Hydrochloride ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Ammonium Bromide ◽  
Molecular Weights ◽  
Viral Particles

Vaccinia virus particles were dissociated into their constituent polypeptides and analysed by sodium dodecyl sulfate (SDS) gel electrophoresis. Thirty-three distinct polypeptide bands were identified and their molecular weights ranged between 11 000 and 150 000 daltons.Specific staining of gels containing polypeptides of dissociated virions revealed the presence of eight glycopeptides. No lipopeptides were detected.Analysis of chemical extracts (urea, guanidine hydrochloride, and alkali treatment) of the virus by SDS gel electrophoresis indicated that a total of 10 to 14 different polypeptides ranging in molecular weights from 11 000 to 70 000 daltons were solubilized.Analysis of detergent extracts and of the remains of extracted viral particles has shown that the detergent Nonidet P-40 (NP-40) solubilized a total of 11 polypeptides of which 6 were glycopeptides. The other detergents sodium deoxycholate (SDC) and cetyl trimethyl ammonium bromide (CTAB) were not as selective, both solubilizing more than 25 of the polypeptides composing the virus. Gel electrophoresis results also indicated that most of the small molecular weight (11 000–70 000 daltons) polypeptides were readily solubilized by NP-40, SDC, and CTAB, while those with molecular weights of 70 000 daltons and higher were not well solubilized.The effects of detergents were also analysed by electron microscopy. Evidence was obtained for subpopulations of viral particles having different susceptibility to detergent extraction.

scholarly journals Interaction of ATP with a Small Heat Shock Protein from Mycobacterium leprae: Effect on Its Structure and Function

2015 ◽  
Vol 9 (3) ◽  
pp. e0003661 ◽  
Author(s):  
Sandip Kumar Nandi ◽  
Ayon Chakraborty ◽  
Alok Kumar Panda ◽  
Sougata Sinha Ray ◽  
Rajiv Kumar Kar ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Small Heat Shock Protein ◽  
Mycobacterium Leprae ◽  
Structure And Function ◽  
And Function

scholarly journals hsp26 of Saccharomyces cerevisiae is related to the superfamily of small heat shock proteins but is without a demonstrable function.

1989 ◽  
Vol 9 (11) ◽  
pp. 5265-5271 ◽  
Author(s):  
R E Susek ◽  
S L Lindquist
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Mutational Analysis ◽  
Small Heat Shock Protein ◽  
Major Effect ◽  
Selfish Dna ◽  
Dna Elements ◽  
Cloned Gene ◽  
Shock Proteins

Analysis of the cloned gene confirms that hsp26 of Saccharomyces cerevisiae is a member of the small heat shock protein superfamily. Previous mutational analysis failed to demonstrate any function for the protein. Further experiments presented here demonstrate that hsp26 has no obvious regulatory role and no major effect on thermotolerance. It is possible that the small heat shock protein genes originated as primitive viral or selfish DNA elements.

scholarly journals The Effect of Oxidized Dopamine on the Structure and Molecular Chaperone Function of the Small Heat-Shock Proteins, αB-Crystallin and Hsp27

2021 ◽  
Vol 22 (7) ◽  
pp. 3700
Author(s):  
Junna Hayashi ◽  
Jennifer Ton ◽  
Sparsh Negi ◽  
Daniel E. K. M. Stephens ◽  
Dean L. Pountney ◽  
...  
Keyword(s):  
Heat Shock ◽  
Protein Aggregation ◽  
Molecular Chaperone ◽  
Cross Linking ◽  
Αb Crystallin ◽  
The Cross ◽  
Shock Proteins ◽  
And Function

Oxidation of the neurotransmitter, dopamine (DA), is a pathological hallmark of Parkinson’s disease (PD). Oxidized DA forms adducts with proteins which can alter their functionality. αB-crystallin and Hsp27 are intracellular, small heat-shock molecular chaperone proteins (sHsps) which form the first line of defense to prevent protein aggregation under conditions of cellular stress. In vitro, the effects of oxidized DA on the structure and function of αB-crystallin and Hsp27 were investigated. Oxidized DA promoted the cross-linking of αB-crystallin and Hsp27 to form well-defined dimer, trimer, tetramer, etc., species, as monitored by SDS-PAGE. Lysine residues were involved in the cross-links. The secondary structure of the sHsps was not altered significantly upon cross-linking with oxidized DA but their oligomeric size was increased. When modified with a molar equivalent of DA, sHsp chaperone functionality was largely retained in preventing both amorphous and amyloid fibrillar aggregation, including fibril formation of mutant (A53T) α-synuclein, a protein whose aggregation is associated with autosomal PD. In the main, higher levels of sHsp modification with DA led to a reduction in chaperone effectiveness. In vivo, DA is sequestered into acidic vesicles to prevent its oxidation and, intracellularly, oxidation is minimized by mM levels of the antioxidant, glutathione. In vitro, acidic pH and glutathione prevented the formation of oxidized DA-induced cross-linking of the sHsps. Oxidized DA-modified αB-crystallin and Hsp27 were not cytotoxic. In a cellular context, retention of significant chaperone functionality by mildly oxidized DA-modified sHsps would contribute to proteostasis by preventing protein aggregation (particularly of α-synuclein) that is associated with PD.

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