Golestan University of Medical Sciences Repository

Self-assembly of tissue transglutaminase into amyloid-like fibrils using physiological concentration of Ca 2+

Kalhor, H.R. and Shahin V, F. and Fouani, M.H. and Hosseinkhani, H. (2011) Self-assembly of tissue transglutaminase into amyloid-like fibrils using physiological concentration of Ca 2+. Langmuir, 27 (17). pp. 10776-10784. ISSN 07437463 (ISSN)

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Abstract

Tissue transglutaminase (tTG or TG2) is a member of the transglutaminase family that catalyzes calcium dependent formation of isopeptide bonds. It has been shown that the expression of TG2 is elevated in neurodegenerative diseases such as Parkinson's, Huntington's, and Alzheimer's. We have investigated the self-assembly of TG2 in vitro. First, using software, hot spots, which are prone for aggregation, were identified in domain 2 of the enzyme. Next we expressed and purified recombinant TG2 and its truncated version that contains only the catalytic domain, and examined their amyloidogenic behavior in various conditions including different temperatures and pHs, in the presence of metal ions and Guanosine triphosphate (GTP).To analyze various stages leading to TG2 fibrillation, we employed various techniques including Thioflavin T (ThT) binding assay, Congo-Red, birefringence, Circular Dichroism (CD), 8-anilino-1-naphthalene sulfonic acid (ANS) binding, Transmission Electron Microscopy (TEM) and Atomic Force Microscopy (AFM). Our results indicated that using low concentrations of Ca 2+, TG2 self-assembled into amyloid-like fibrils; this self-assembly occurred at the physiological temperature (37 °C) and at a higher temperature (57 °C). The truncated version of TG2 (domain 2) also forms amyloid-like fibrils only in the presence of Ca 2+. Because amyloid formation has occurred with domain 2 alone where no enzymatic activity was shown, self-cross-linking by the enzyme was ruled out as a mechanism of amyloid induction. The self-assembly of TG2 was not significant with magnesium and zinc ions, indicating specificity of the self-assembly for calcium ions. The calcium role in self-assembly of TG2 into amyloid may be extended to other proteins with similar biophysical properties to produce novel biomaterials. © 2011 American Chemical Society.

Item Type: Article
Additional Information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Langmuir [Field not mapped to EPrints] C2 - 21790128 [Field not mapped to EPrints] AD - Department of Molecular Medicine, School of Advanced Technology for Medical Sciences, Golestan University of Medical Sciences, Gorgan, Iran [Field not mapped to EPrints] AD - Nanomedicine and Tissue Engineering Research Center, Taleghani Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran [Field not mapped to EPrints] AD - Departments of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran [Field not mapped to EPrints] AD - Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology, Taipei, 10607, Taiwan [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled Keywords: Alzheimer, Amyloid formation, Amyloid-like fibril, Binding assays, Biophysical properties, Calcium dependent, Calcium ions, Catalytic domains, Congo-red, Enzymatic activities, Guanosine triphosphate, Higher temperatures, Hot spot, In-vitro, Isopeptide bond, Low concentrations, Physiological concentrations, Physiological temperature, Self-assembled, Sulfonic acid, Thioflavin T, Tissue transglutaminase, Transglutaminases, Truncated versions, Zinc ions, Atomic force microscopy, Calcium, Dichroism, Enzymes, Glycoproteins, Magnesium, Metal ions, Naphthalene, Neurodegenerative diseases, Physiology, Self assembly, Tissue, Transmission electron microscopy, Proteins, amyloid, calcium, protein glutamine gamma glutamyltransferase, recombinant protein, tissue transglutaminase 2, human, article, biocatalysis, chemical structure, chemistry, dose response, drug effect, erythrocyte, human, isolation and purification, metabolism, protein conformation, temperature, Amyloid, Biocatalysis, Calcium, Dose-Response Relationship, Drug, Erythrocytes, Humans, Models, Molecular, Protein Conformation, Recombinant Proteins, Temperature, Transglutaminases
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
موارد کلی
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 19 Apr 2015 05:21
Last Modified: 16 May 2015 06:35
URI: http://eprints.goums.ac.ir/id/eprint/2096

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