Alpha crystallin domain mutations increase holdase capacity of the archeal small heat shock protein, Tpv HSP 14.3

Date

2016-07-25

Author

Kocabıyık, Semra

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

189
views

0
downloads

Protein stability involves the ability of native, folded protein structure to withstand the disruptive, denaturing effects of the external environment. Proteins used in many medical or biotechnological processes are often subjected to conditions which may trigger their degradation, aggregation or inactivation. Therefore, stabilization of proteins is one of the most challenging tasks that has to be addressed in protein based technologies. Many methods have been proposed for improvement of chemical and/or physical instability of proteins. An effective strategy is exploiting the capacity of molecular chaperone sHSPs that act as holdases to bind a wide range of proteins and thereby impede the deleterious effects of their aggregation. This current approach has been promising for various successful applications including proteomics, nanobiotechnology, bioproduction, bioseparation. The focus of our research is the use of an archeal sHSP (Tpv HSP14.3) to enhance thermal stability of a mesophilic enzyme citrate synthase (opt. temp 35°C) as the model substrate. Temperature dependent chaperone activities of the wild type sHSP and its mutant variant TpvHSP14.3 (with ACD mutations) were investigated, comparatively. Thermal stability of the citrate synthase at 47°C was increased significantly by wild type and mutant sHSP as revealed by 9-fold and 11-fold increase in its activity, respectively. Structural analysis showed that increased chaperone activity displayed by the mutant sHSP may be due to local structural change in the β6–β7 zone of ACD resulting in perturbation of oligomer integrity which then impacts protein binding capacity.

Subject Keywords

Biochemistry, Molecular Biology

URI

https://hdl.handle.net/11511/46694

Journal

NEW BIOTECHNOLOGY

DOI

https://doi.org/10.1016/j.nbt.2016.06.1083

Collections

Department of Biology, Article

Suggestions

OpenMETU
Core

Immunotherapeutic applications of CpG ODN Gürsel, Mayda (2006-06-01) Bacterial DNA and synthetic oligodeoxynucleotides (ODN) expressing unmethylated CpG motifs stimulate the mammalian immune system to mount a rapid innate immune response. This response is characterized by the production of polyreactive IgM, immunomodulatory cytokines and chemokines. CpG ODN directly stimulate lymphocytes, natural killer cells and professional antigen-presenting cells (such as macrophages and dendritic cells). Owing to the strength and nature of this stimulation, CpG ODN are being harnessed f...
Proton relay network in P450cam formed upon docking of putidaredoxin Ugur, Ilke; Chandrasekhar, Prasanna (Wiley, 2020-04-01) Cytochromes P450 are versatile heme-based enzymes responsible for vital life processes. Of these, P450cam (substrate camphor) has been most studied. Despite this, precise mechanisms of the key OO cleavage step remain partly elusive to date; effects observed in various enzyme mutants remain partly unexplained. We have carried out extended (to 1000 ns) MM-MD and follow-on quantum mechanics/molecular mechanics computations, both on the well-studied FeOO state and on Cpd(0) (compound 0). Our simulations include...
The NF-kappa B target genes ICAM-1 and VCAM-1 are differentially regulated during spontaneous differentiation of Caco-2 cells Astarci, Erhan; Sade, Asli; Cimen, Ismail; SAVAŞ, BERNA; Banerjee, Sreeparna (Wiley, 2012-08-01) Intestinal epithelial differentiation entails the formation of highly specialized cells with specific absorptive, secretory, digestive and immune functions. Cellcell and cellmicroenvironment interactions appear to be crucial in determining the outcome of the differentiation process. Using the Caco-2 cell line, which undergoes spontaneous re-differentiation when grown past confluency, we observed a loss of VCAM-1 (vascular cell adhesion molecule 1) mRNA expression, while ICAM-1 (intercellular cell adhesion m...
Combinatorial Tau pseudophosphorylation: markedly different regulatory effects on microtubule assembly and dynamic instability than the sum of the individual parts. Kiriş, Erkan; Sargin, ME; Gaylord, MR; Altinok, A; Rose, K; Manjunath, BS; Jordan, MA; Wilson, L; Feinstein, SC (American Society for Biochemistry & Molecular Biology (ASBMB), 2011-04-22) Tau is a multiply phosphorylated protein that is essential for the development and maintenance of the nervous system. Errors in Tau action are associated with Alzheimer disease and related dementias. A huge literature has led to the widely held notion that aberrant Tau hyperphosphorylation is central to these disorders. Unfortunately, our mechanistic understanding of the functional effects of combinatorial Tau phosphorylation remains minimal. Here, we generated four singly pseudophosphorylated Tau proteins ...
Structural Insights into Alternate Aggregated Prion Protein Forms POLANO, maurizio; Bek, Alpan; BENETTİ, federico; lazzarino, marco; LEGNAME, giuseppe (Elsevier BV, 2009-11-13) The conversion of the cellular form of the prion protein (PrPC) to an abnormal, alternatively folded isoform (PrPSc) is the central event in prion diseases or transmissible spongiform encephalopathies. Recent studies have demonstrated de novo generation of murine prions from recombinant prion protein (recPrP) after inoculation into transgenic and wild-type mice. These so-called synthetic prions lead to novel prion diseases with unique neuropathological and biochemical features. Moreover, the use of recPrP i...

Citation Formats

S. Kocabıyık, “Alpha crystallin domain mutations increase holdase capacity of the archeal small heat shock protein, Tpv HSP 14.3,” NEW BIOTECHNOLOGY, pp. 0–0, 2016, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/46694.