... formation of huntingtin-containing inclusion bodies in Huntington’s disease encourages cell survival, whereas monomers and small inclusion bodies of huntingtin, a protein possessing abnormal polyQ ... inclusions develop in cultured nonmuscle cells deficient in desmin, again demonstrating inclusion body construction independent of intermediate fialments [62] Interestingly, inclusion body formation ... lines decreases invasiveness, reduces matrix metalloproteinases in vitro and eliminates production of avb3 integrin, a protein missing in normal melanocytes but often manufactured during the invasive...
... aB-crystallin uninduced at the 0, and h time points Lane contains cell homogenates of aB-crystallin not induced Lanes 8–10 contain aB-crystallin induced with IPTG at the 0, and h time points The cells ... 2–5 show protein expression in cells containing the pET 16b vector alone at selected times from to h Lane is the protein expression in cells containing the pET 16b vector alone not induced with ... between the two proteins The conserved core a-crystallin domain observed in proteins of the sHsp spans residues of E67–I161 inthe aB-crystallin sequence are consistent with previous studies on the...
... were mainly interested inthe irreversible changes inthe structure of G-actin that were induced by heating Therefore in all experiments the samples of actin in G-buffer were heated for h at the ... with intact actin Fig Effect of heating on the kinetics of polymerization (A) and saltinduced increase of the light scattering (B) of actin Actin (15 lM) containing 10% or pyrene-labeled actin in ... actin inthe sample Results Heat denaturation of actin Before starting the investigation of the HSP25–actin interaction it was desirable to characterize the properties of intact and heated actin...
... a-crystallin domain In contrast they show only 41% identity and 50% similarity inthe N-terminal arm, suggesting substrate specificity is determined by the N-terminal arm The N-terminus of PsHsp18.1.C-I ... containing the R120G mutation linked to desmin-related myopathy One of the authors’ interpretations for the effect was a possible change inthe availability of substrate binding sites resulting in a ... Residues inthe N-terminal region shown in bold correspond to sequences conserved in all class I or class II proteins, respectively Residues inthe C-terminus in bold correspond to the conserved...
... wild-type proteins The corresponding glycine residues in wheat Hsp16.9 and M jannaschii Hsp16.5 are located in a short connecting loop, between b-strands and 9, which is involved in dimer interaction ... chaperone activity Thus, it was speculated that G114 might be involved in substrate binding [27] Inthe present study, we investigated thein uence of temperature and concentration on sHsp complexes ... according to the manufacturer’s instructions The correct nucleotide sequence was confirmed by automated DNA sequencing All sHsps used in this study carry a C-terminal hexahistidine tag Protein expression...
... expressing the Tet repressor (Invitrogen) were transfected with pcDNA4/TO (Invitrogen) containing the coding sequences for wild type aB-crystallin, aB-crystallin S19D/S45D/S59D or aB-crystallin S19A/S45A/S59A ... that in all cells the cytoplasmic staining was strongly reduced Only cells expressing aB-crystallin S19D/S45D/S59D show the nuclear bodies, indicating that at least part of the detergentinsoluble ... treatment; only the staining of the nucleoli remained, indicating that most of the DNA was digested However aB-crystallin could still be detected in SC35 speckles (Fig 3B, a and c) The RNase-treated...
... protein indicates that the structure of the N-terminal domain is maintained, at least inthe vicinity of the tryptophan residues The increase in exposed clustered hydrophobicity observed for the ... that the binding of target proteins occurs inthe groove between monomers, and involves a b-sheet region located at the beginning of the a-crystallin domain corresponding to residues 70–88 in aA-crystallin ... facilitate interaction between the extension and hydrophobic chaperone binding sites, resulting inthe binding sites being less accessible to the target protein, leading to a decrease in target protein...
... [17,41,43–46] a-crystallin domain of p26 The p26 a-crystallin domain is similar in sequence to this region in other sHSPs, including wheat Hsp16.9; the protein confers thermotolerance on transformed ... conserved in many other sHSPs, including those inthe sequence 110REFRRRY116, where substitutions were generated Examination of mutations within the selected sequence indicated that b-strand is involved ... for h in either the absence or presence of p26, and then enzyme activity was determined p26 concentrations are indicated and the p26 variants are inthe same order in each histogram group The standard...
... (where Ao and At are the intensities of the band of intact protein at the beginning of trypsinolysis and at the fixed time of trypsinolysis) against the time of incubation (Fig 8D) The apparent rate ... observed inthe case of the K137E mutant of HSP22 (Fig 4B); however, in this case, the intensity of the band corresponding to the HSP22 dimer was more intense than inthe case of the wild-type protein ... proteins According to the predictions, K137 is located either inthe C-terminal part of the b5–b7 loop or inthe N-terminal part of the b7 strand, whereas K141 is located inside the b7 strand...
... However, another report showed that the modification rather hinders the DNAbinding activity of HSF2, without interfering with its trimerization [97] A subsequent study further strengthened the molecular ... [102] Inthe olfactory epithelium of 3–6-week-old mice, the expression profile of HSF1 remains constant However, a significant increase inthe DNA-binding activity of HSF1 can be detected during the ... HSF1 inthe olfactory epithelium warrants further investigations In accordance with the fundamental role of HSF1 inthe heat shock response, the requirement of HSF1 and HSF4 in development of the...
... performed to analyze the EGFP-containing inclusions Figure 1F shows that the expression of Hsp70 together with Hdj-1 (Hsp40) did decrease the average size of the EGFP-containing inclusion bodies (average ... position of the more intensively oxidized polypeptide inthe assay The bracket underlines the domain (molecular mass range 10–40 kDa) of the oxyblots that contains the greatest changes in protein oxidation ... of the initiator ATG in exon (Ex1) of the 67 exon-containing htt gene [5] Pathogenesis in HD correlates with the cleavage of mutated htt and the release of an N-terminal fragment bearing the...
... shock/a-crystallin proteins aid construction of oligomers from smaller building blocks arising by interactions between residues within a-crystallin domains [7,8,21,62] Contrary to other reports, the C-terminal ... proteins in vitro [12] These polypeptides are unlikely to oligomerize, either in vivo or in vitro Clearly, our data strengthen the notion that small heat shock/a-crystallin proteins function in ... the latter modification was small In contrast, loss of 42 residues from the N-terminus of a rice small heat shock/a-crystallin protein increased oligomer size [23] Deleting the last 16 C-terminal...
... of increasing quantities of intact wild-type Hsp20 results in an increase of the lag period and a decrease inthe amplitude of light scattering Significant retardation of the insulin B-chain aggregation ... was higher than that of the intact wild-type Hsp20 (Fig 1B) By analyzing the distribution of the charge residues inthe C-terminal region of Hsp20, and by calculating the theoretical pI values ... a-crystallin At pH 7.0, the heating of isolated ADH inthe absence of divalent cations was accompanied by aggregation and a large increase inthe light scattering (Fig 7, curve 1) Addition of increasing...
... Other conditions were the same as in Fig 1A The Rh values were determined from the Rh distributions obtained after F-actin thermal denaturation at 70 °C The Hsp27-3D ⁄ actin weight ratios inthe ... determining the quantity of free Hsp27-3D, we can calculate the amount of Hsp27-3D involved inthe complexes with denatured actin, and by this means estimate the stoichiometry Hsp27-3D ⁄ actin in these ... 100 kDa), contained isolated Hsp27-3D The size of this peak was clearly increased with increasing initial concentrations of Hsp27-3D inthe incubation mixture Thus, knowing the initial total concentration...
... 153-IVPITP-158 of the p26 C-terminal extension are included inthe model and within this sequence 5236 p26 contains two novel N-terminal sequences, one enriched in glycine and the other in arginine Additionally, ... motifs inthe p26 N-terminal extension and another inthe C-terminus The deletion of these motifs, termed G (multiple glycine), R (multiple arginine), and TS (multiple threonine ⁄ serine), was ... oligomerization, but deleting the entire C-terminal extension, including the TS sequence and the conserved I ⁄ V-XI ⁄ V motif (154-VPI-156 in p26) had more impact [11] Eliminating the I ⁄ V-X-I ⁄ V motif...
... encoding the isolated a-crystallin domain or the a-crystallin domain including either the N-terminal region or the C-terminal extension were constructed in a similar manner Mutations leading to single ... shown (S) consisting of the a-crystallin domain alone [HspH(a)], the a-crystallin domain plus the N-terminal region [HspH(Na)], or the a-crystallin domain followed by the C-terminal extension [HspH(aC)], ... constructed, in which the isoleucines were replaced by alanine, either individually, resulting in HspH(I133A) and HspH(I135A), or simultaneously, leading to HspH(II133,135AA) Finally, we searched for the...
... the injurious agent and the injured tissue The inflammatory response to infections as well as other stimuli involves a myriad of defenses, including both the innate and adaptive arms of the immune ... spirochete inthe stomach of dogs This finding has since been verified by numerous scientists including Salomon in 1896 and Krienitz in 1906 who related a similar finding to gastric cancer in a human ... and opinions exist inthe field [49-51] The innate immune responses induced by HSPs include cytokine and chemokine release by professional APCs and T-cells, maturation of DCs by upregulating the...
... demonstrated the induction of HSPs 72 in RGCs, the whole body hyperthermia was found to excessively induce HSPs 72 inthe entire body causing to overexpression inthe immune system, and the intraperitoneal ... enhance the induction efficiency of local HSPs 72 are also presented In addition, a newly designed infusion technique to inject the SPNPs into the retina layer will be presented Finally, chapter concludes ... proteins that exist in all of the living creatures covering bacteria to human beings HSPs can be induced in living cells by applying various stresses such as thermal stress (hyperthermia), metabolic...
... contamination is being highlighted recently in assessing the role of HSP in innate immunity [Bausinger et al., 2002a] In fact, several studies have shown that endotoxin-free HSPs are not able to induce ... al., 2001; Wendling et al., 2000] In addition, HSP can suppress the production of interleukin-4 (IL-4) and interleukin-5 (IL-5), while concurrently increasing the production of IL-10 in a mouse model ... of thein vitro cell culture system to investigate the role of the HSPs in antigen presentation and signaling Furthermore, we examine the immunogenicity and regulatory properties of HSPs using...
... of HSF-1 DNA binding inthe heart [26] The reasons for the different response of dipterans and mammals to PARP-1 inhibition are unknown It is plausible that factors including signaling mediators ... prevents their binding to DNA On the contrary, inhibition of PARP-1 enables the binding of the transcription factors to their specific DNA sites [5] Thus, it is possible that both physical interaction ... repressing factor of the heat shock response by regulating the expression of HSP-70 Both protein-protein interaction and catalytic activity of the PARP protein play a key role in modulation of the...