is targeting of clu to the nucleus an experimental artifact
Tài liệu Báo cáo khoa học: The small heat shock proteins and their role in human disease pptx
... and disease more limited than for other chaperones, but this is changing as the application of genomics and proteomics reveals sHSP characteristics and their medical importance emerges In this ... disease Many diseases are associated with changes to sHSPs occurring either as a result of mutation or by post-translational changes, and these are outlined below and described in the text of ... proteins and disease as does the R116C mutation However, in contrast to R116C aA-crystallin, the R49C variant localizes to the cell nucleus and the cytoplasm, superficially suggesting a relationship to...
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Báo cáo khoa học: Functional similarities between the small heat shock proteins Mycobacterium tuberculosis HSP 16.3 and human aB-crystallin docx
... vitro The molecular chaperone activity of recombinant MTB HSP 16.3 was enhanced in the presence of ATP which is consistent with previous findings of the effect of ATP on recombinant human aB-crystallin ... Figure is the SDS/PAGE and Western immunoblot analyses of the expression and purification of MTB HSP 16.3 Induction of protein expression with IPTG resulted in the appearance of a protein band at ... 1A, lanes and 3) The expressed MTB HSP 16.3 was purified by a combination of anion exchange, hydrophobic interaction and size exclusion chromatography (lanes 4–6) Western immunoblot analysis of...
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Tài liệu Báo cáo khoa học: Interaction of the small heat shock protein with molecular mass 25 kDa (hsp25) with actin doc
... composition of both supernatant and pellet was determined by quantitative SDS gel electrophoresis The quantity of actin in the pellet was plotted against the total quantity of actin in the sample ... effect on the extent of polymerization of intact actin, whereas both 2D and 3D mutants decreased the quantity of polymerized actin in the pellet It is worthwhile to mention that the mutants of HSP25 ... was added and after mixing the samples were immediately subjected to ultracentrifugation The quantity of actin in the pellet is plotted against the total quantity of actin in the sample The results...
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Báo cáo khóa học: Chaperone activity of cytosolic small heat shock proteins from wheat pptx
... Sedimentation velocity analysis of TaHsp16.9C-I and TaHsp17.8C-II Shown is the integral distribution plot from the van Holde–Weischet analysis of the data obtained for TaHsp16.9C-I (s) and TaHsp17.8C-II ... points and error bars reflect the mean and standard deviation of three replicates Results Comparison of TaHsp16.9 C-I and TaHsp17.8 C-II To produce recombinant wheat class I and II sHSPs for these ... absence of the sHsp This may reflect the formation of very large aggregates of MDH that also include the sHsp, as evidenced by the loss of sHsp from the SEC profile under these conditions At this low...
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Báo cáo khoa học: Temperature and concentration-controlled dynamics of rhizobial small heat shock proteins doc
... indicated by the dotted lines assemblies A transfer of CS and HspH into the void volume, at the expense of the other fractions, was confirmed by SDS-PAGE (Fig 7C) Neither HspH nor CS shifted into the void ... inspection of the dynamic properties of this protein and an equivalent mutant in HspB, revealed that these variants form labile complexes that dissociate much more readily than the wild-type proteins The ... between 20 °C and 31 °C The most abundant species was adjusted to 100% peak intensity (Inset) Graph of the ratio of summed peak areas for charge state series of the 12mer relative to the dissociated...
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Báo cáo khoa học: Mimicking phosphorylation of the small heat-shock protein aB-crystallin recruits the F-box protein FBX4 to nuclear SC35 speckles docx
... Hasegawa, K., Goto, S., Ito, H & Kato, K (1995) Induction of the synthesis of hsp27 and aB-crystallin in tissues of heat-stressed rats and its suppression by ethanol or an a1-adrenergic antagonist J Biochem ... cultures of lens epithelial cells derived from wild type and aB–/– mouse lenses [37] to assess the specificity of the K79 antiserum Our analysis showed that this antibody diffusely stained the cytoplasm ... immunocytochemical analysis, the antibody was added undiluted to the fixed cells In addition another monoclonal antibody to aB-crystallin (2D2B6) [35], and a polyclonal peptide antibody to the N-terminal...
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Báo cáo khoa học: Glutamic acid residues in the C-terminal extension of small heat shock protein 25 are critical for structural and functional integrity pptx
... prepared and included as a control The role of the C-terminal extension and each of the mutated residues was investigated by comparison of the structure and function of these mutants with those of ... performed on wild-type and mutant Hsp25 to detect any changes in the environment of the tryptophan residues resulting from the mutations The tryptophan residues of Hsp25 are located in the N-terminal ... that the C-terminal extension is involved in the quaternary structural arrangement of sHsps 5886 The alteration of the properties of the C-terminal extension also leads to significant changes to the...
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Báo cáo khoa học: Structural and functional roles for b-strand 7 in the a-crystallin domain of p26, a polydisperse small heat shock protein from Artemia franciscana pdf
... state of metabolic arrest, termed quiescence, they survive for several years without oxygen This is arguably the ultimate indifference to anoxia of any metazoan [42], and qualifies the organism, ... goat anti-rabbit IgG The top of each gradient is to the right and fractions are numbered across the top The positions of the molecular mass markers alpha-lactalbumin, 14.2 kDa; carbonic anhydrase, ... result of the residue’s location at the edge of the dimer–dimer interface The maximum oligomer size obtained with p26 R114A was 500 kDa, closer to the mass of the WT oligomer than the other variants...
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Báo cáo khoa học: Effect of mutations in the b5–b7 loop on the structure and properties of human small heat shock protein HSP22 (HspB8, H11) pptx
... substrates to sHsp [17] The importance of this part of molecule of the sHsp is supported by the fact that mutations in the loop connecting the b5 and b7 strands, or in the b7 strand of sHsp, often ... study analyses the structure and properties of K137E and the K137,141E mutant of human HSP22, aiming to provide new information on the structure of sHsp and to shed new light on their role in the ... sHsp The primary structure of the a-crystallin domain of human sHsp is very conservative and the loop connecting the b5 and b7 strands is shorter than the corresponding loop connecting the b5 and...
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Tài liệu Báo cáo khoa học: Regulation of the members of the mammalian heat shock factor family doc
... monomer -to- trimer transition of HSF1 in the olfactory epithelium warrants further investigations In accordance with the fundamental role of HSF1 in the heat shock response, the requirement of HSF1 and ... [6,7,10,15,113–115] In spermatogenesis, disruption of both Hsf1 and Hsf2 leads to a more pronounced phenotype (i.e male sterility) than disruption of either factor alone The phenotype of the double knockout ... Bjork and L Sistonen ¨ Pirkkala L, Nykanen P & Sistonen L (2001) Roles of ¨ the heat shock transcription factors in regulation of the heat shock response and beyond FASEB J 15, 1118–1131 Fujimoto...
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Tài liệu Báo cáo khoa học: Analysis of oxidative events induced by expanded polyglutamine huntingtin exon 1 that are differentially restored by expression of heat shock proteins or treatment with an antioxidant ppt
... cases (72Q and 103Q), a broad distribution of the size of the inclusions was noticed Moreover, the percentage of cells presenting inclusions as well as the distribution of the size of the inclusions ... Note the presence of the granules in the cytoplasm of the cells Bar, 20 lm (D) The percentage of EGFP-containing cells displaying granules is shown The average percentages, including standard ... was found to correlate with the number of CAG repeats in the httEx1-polyQ polypeptide [15] The mechanism responsible for the appearance of an oxidative stress in response to the presence of aggregated...
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Tài liệu Báo cáo Y học: Functional analysis of a small heat shock/a-crystallin protein from Artemia franciscana docx
... polyacrylamide gels, transferred to nitrocellulose, and reacted with antibody to the His6 epitope tag The top of each gradient is to the left of the figure, and numbers across the top indicate successive ... polyacrylamide gels, transferred to nitrocellulose, and reacted with antibody to p26 using the ECL procedure The top of each gradient is to the left of the figure, and numbers across the top indicate successive ... 30 °C and induced with IPTG were probed with antibody to p26 Films were scanned and absorbance of the p26 band in each lane, in arbitrary units, determined The amounts of sample applied to the...
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Báo cáo khóa học: Some properties of human small heat shock protein Hsp20 (HspB6) potx
... Rearrangement of the complexes formed by the 3D mutant of Hsp27 and the wild-type Hsp20 A mathematical summation of the elution profiles of the isolated 3D mutant of Hsp27 and isolated wild-type Hsp20 is ... results in the dissociation of small oligomers of Hsp20 and its S16D mutant to monomers that tend to unfold and aggregate These aggregates are retarded on the top of the column and therefore not ... electrophoresis, the samples were either treated with an excess of b-mercaptoethanol or loaded onto the gel in the absence of b-mercaptoethanol In the second method, Hsp20 (0.75 mgÆmL)1) in 0.2 M triethanolamine...
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Báo cáo khoa học: Small heat shock protein Hsp27 prevents heat-induced aggregation of F-actin by forming soluble complexes with denatured actin docx
... hundreds and even thousands of actin subunits This is consistent with a recently proposed dissociative mechanism of F-actin thermal denaturation [1] One of the main features of this mechanism is that ... where D is the diffusion coefficient obtained from the DLS measurements, kB is Boltzmann’s constant, T is the absolute temperature, and g is the shear viscosity of the solvent The viscosity of the ... proteins is dangerous for the cell, and this is especially important in the case of abundant proteins, such as actin There are different mechanisms for preventing formation of insoluble aggregates, and...
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Báo cáo khoa học: Characterization of novel sequence motifs within N- and C-terminal extensions of p26, a small heat shock protein from Artemia franciscana potx
... blotted to nitrocellulose and reacted with antibody to p26 followed by HRPconjugated goat antirabbit IgG The top of each gradient is to the right and fractions are numbered across the top The molecular ... Council of Canada Discovery Grant, a Nova Scotia Health Research Foundation ⁄ Canadian Institutes of Health Regional Partnership Plan Grant, and a Heart and Stroke Foundation of Nova Scotia Grant to ... and TS were similar to each other (P > 0.05) and significantly higher than the thermotolerance conferred by variant R (P < 0.05) However, because the amount of TS p26 in transformed bacteria was...
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Báo cáo Y học: A critical motif for oligomerization and chaperone activity of bacterial a-heat shock proteins pot
... as one of the three main consensus regions of the a-Hsp family [5] There is little experimental evidence to circumstantiate the role of the C-terminal extension, and even less concerning the conserved ... integrity of the I-X-I motif is critical for assembly of functional a-Hsp proteins, and that the isolated a-crystallin domain is unable to dimerize EXPERIMENTAL PROCEDURES with NdeI and XhoI and ligated ... introduced to HspH and HspF, and asterisks indicate alanine exchange mutations in HspH Note that four N-terminal residues were included in HspH(a) and HspH(aC) to avoid disturbance of potential...
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báo cáo khoa học: "Heat-shock proteins in infection-mediated inflammation-induced tumorigenesis" potx
... Carcinoma on the Floor of the Pelvis: Two Discoveries in Cancerous Disease Transactions of the American Microscopial Sciety 1898, 20:165-176 Balkwill F, Mantovani A: Inflammation and cancer: back to Virchow? ... relative to the diversity of human neoplasms and pathogens Nevertheless, the worldwide population burden of infectious organisms makes understanding their role in human disease of paramount importance ... HSP27 of the inducible small HSP family has been shown to inhibit the mitochondrial release of SMAC (second mitochondrial-derived activator of caspase), the master regulator of apoptosis, to confer...
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Local induction of heat shock proteins using magnetic fluid hyperthermia for ocular neuroprotection in glaucoma
... INTRODUCTION 1.3 Organization of Thesis Chapter presents the background, motivations, and objectives of this thesis Chapter reviews the previous works relevant to this project and discusses the theoretical ... ferrofluids can be used in an in-vivo environment to increase the temperature of tissues or organs to induce HSPs or to treat cancers without the assistance of other therapy methods Since then, ... technique to introduce SPNPs to the surface of retina layer and the histological exam results to investigate the distribution status of the injected SPNPs and cell apoptosis: (a) Injection of uncoated...
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Immunomodulatory roles of heat shock proteins
... assistance and comforting words To all the members and colleagues in the lab, for accompanying me through out the course of my study To Dr Leslie, Jeremy, James and Sean from the AHU, and Ms Huang ... Huang C-H from Prof Chua KY’s lab, for their help in the animal studies To Karen, Weiping and Huang Bo, for their assistance in DNA sequencing To Dr Lu and Jason, for their help and discussion in ... Belli et al., 2002] The mechanism of HSP-peptide-induced immunity is proposed to happen in the following manner: HSP mediates the transfer of exogenous peptides onto the major histocompatibility...
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Báo cáo y học: "Small interfering RNA mediated Poly (ADP-ribose) Polymerase-1 inhibition upregulates the heat shock response in a murine fibroblast cell line" pptx
... to wild type fibroblasts [27] In gene knockout cell lines, unexpected compensatory or redundant mechanisms develop in response to the missing gene and can confound experimental observations To ... Immunoreaction was visualized by chemiluminescence Data analysis All values in the figures and text are expressed as mean ± SEM The results were examined by analysis of variance followed by the Bonferroni’s ... is plausible that factors including signaling mediators have evolved different effector proteins that can affect the heat shock response and its regulation by PARP-1 Another potential mechanism...
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