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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Recombinant expression, purification and preliminary biophysical and structural studies of C-terminal carbohydrate recognition domain from human galectin-4

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Author(s):
Rustiguel, Joane K. [1] ; Kumagai, Patricia S. [2] ; Dias-Baruffi, Marcelo [3] ; Costa-Filho, Antonio J. [4] ; Nonato, Maria Cristina [1]
Total Authors: 5
Affiliation:
[1] Univ Sao Paulo, Lab Cristalog Prot, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP - Brazil
[2] Univ Sao Paulo, Inst Fis Sao Carlos, Grp Biofis Mol Sergio Mascarenhas, BR-13563120 Sao Carlos, SP - Brazil
[3] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Lab Glicoimunol, BR-14040903 Ribeirao Preto, SP - Brazil
[4] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Lab Biofis Mol, BR-14040901 Ribeirao Preto, SP - Brazil
Total Affiliations: 4
Document type: Journal article
Source: Protein Expression and Purification; v. 118, p. 39-48, FEB 2016.
Web of Science Citations: 3
Abstract

Galectin-4 (Gal4), a tandem-repeat type galectin, is expressed in healthy epithelium of the gastrointestinal tract. Altered levels of Gal4 expression are associated with different types of cancer, suggesting its usage as a diagnostic marker as well as target for drug development. The functional data available for this class of proteins suggest that the wide spectrum of cellular activities reported for Gal4 relies on distinct glycan specificity and structural characteristics of its two carbohydrate recognition domains. In the present work, two independent constructs for recombinant expression of the C-terminal domain of human galectin-4 (hGal4-CRD2) were developed. His(6)-tagged and untagged recombinant proteins were overexpressed in Escherichia coli, and purified by affinity chromatography followed by gel filtration. Correct folding and activity of hGal4-CRD2 were assessed by circular dichroism and fluorescence spectroscopies, respectively. Diffraction quality crystals were obtained by vapor-diffusion sitting drop setup and the crystal structure of CRD2 was solved by molecular replacement techniques at 1.78 angstrom resolution. Our work describes the development of important experimental tools that will allow further studies in order to correlate structure and binding properties of hGal4-CRD2 and human galectin-4 functional activities. (C) 2015 Elsevier Inc. All rights reserved. (AU)

FAPESP's process: 11/21811-1 - Structural characterization and evaluation of funciontal aspects of tanden-repeat human galectins
Grantee:Maria Cristina Nonato
Support Opportunities: Regular Research Grants
FAPESP's process: 11/21767-2 - Molecular interactions in the mechanism of action of human galectin-4
Grantee:Patricia Suemy Kumagai
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 10/17662-8 - Electron magnetic resonance in studies of structure, function and interactions of biologically-relevant molecules
Grantee:Antonio José da Costa Filho
Support Opportunities: Regular Research Grants
FAPESP's process: 10/16153-2 - Structural characterization and evaluation of functional aspects of human galectins from tandem-repeat group
Grantee:Joane Kathelen Rustiguel Bonalumi
Support Opportunities: Scholarships in Brazil - Doctorate