Low-Resolution Structure and Fluorescence Anisotropy Analysis of Protein Tyrosine Phosphatase η Catalytic Domain
| العنوان: | Low-Resolution Structure and Fluorescence Anisotropy Analysis of Protein Tyrosine Phosphatase η Catalytic Domain |
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| المؤلفون: | Mario de Oliveira Neto, Igor Polikarpov, Rodolfo Iuliano, Huita C. Matozo, Lucas Bleicher, Maria A.M. Santos, Luís Maurício T.R. Lima, Alfredo Fusco |
| المساهمون: | Hc, Matozo, Ma, Santo, M., DE OLIVEIRA NETO, L., Bleicher, Lm, Lima, R., Iuliano, Fusco, Alfredo, I., Polikarpov |
| المصدر: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| بيانات النشر: | Biophysical Society, 2007. |
| سنة النشر: | 2007 |
| مصطلحات موضوعية: | Models, Molecular, Protein Conformation, Dimer, Biophysics, Protein tyrosine phosphatase, Catalysis, chemistry.chemical_compound, Protein structure, rat protein tyrosine phosphatase, Computer Simulation, small angle x ray scattering, Crystallography, Small-angle X-ray scattering, Chemistry, DEP-1, Receptor-Like Protein Tyrosine Phosphatases, Class 3, Proteins, Transmembrane protein, ENZIMAS, Protein Structure, Tertiary, Dissociation constant, Models, Chemical, Anisotropy, Protein Tyrosine Phosphatases, Rat Protein, Fluorescence anisotropy |
| الوصف: | The rat protein tyrosine phosphatase eta, rPTPeta, is a class I "classical" transmembrane RPTP, with an intracellular portion composed of a unique catalytic region. The rPTPeta and the human homolog DEP-1 are downregulated in rat and human neoplastic cells, respectively. However, the malignant phenotype is reverted after exogenous reconstitution of rPTPeta, suggesting that its function restoration could be an important tool for gene therapy of human cancers. Using small-angle x-ray scattering (SAXS) and biophysical techniques, we characterized the intracellular catalytic domain of rat protein tyrosine phosphatase eta (rPTPetaCD) in solution. The protein forms dimers in solution as confirmed by SAXS data analysis. The SAXS data also indicated that rPTPetaCD dimers are elongated and have an average radius of gyration of 2.65 nm and a D(max) of 8.5 nm. To further study the rPTPetaCD conformation in solution, we built rPTPetaCD homology models using as scaffolds the crystallographic structures of RPTPalpha-D1 and RPTPmicro-D1 dimers. These models were, then, superimposed onto ab initio low-resolution SAXS structures. The structural comparisons and sequence alignment analysis of the putative dimerization interfaces provide support to the notion that the rPTPetaCD dimer architecture is more closely related to the crystal structure of autoinhibitory RPTPalpha-D1 dimer than to the dimeric arrangement exemplified by RPTPmicro-D1. Finally, the characterization of rPTPetaCD by fluorescence anisotropy measurements demonstrates that the dimer dissociation is concentration dependent with a dissociation constant of 21.6 +/- 2.0 microM. |
| وصف الملف: | STAMPA |
| اللغة: | English |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4d639b1361d666bf22149769fc089da9Test https://europepmc.org/articles/PMC1877755Test/ |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....4d639b1361d666bf22149769fc089da9 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |