Expression, purification and molecular analysis of the human ZNF706 protein.
BACKGROUND
The ZNF706 gene encodes a protein that belongs to the zinc finger family of proteins and was found to be highly expressed in laryngeal cancer, making the structure and function of ZNF706 worthy of investigation. In this study, we expressed and purified recombinant human ZNF706 that was suitable for structural analysis in Escherichia coli BL21(DH3).


RESULTS
ZNF706 mRNA was extracted from a larynx tissue sample, and cDNA was ligated into a cloning vector using the TOPO method. ZNF706 protein was expressed according to the E. coli expression system procedures and was purified using a nickel-affinity column. The structural qualities of recombinant ZNF706 and quantification alpha, beta sheet, and other structures were obtained by spectroscopy of circular dichroism. ZNF706's structural modeling showed that it is composed of α-helices (28.3%), β-strands (19.4%), and turns (20.9%), in agreement with the spectral data from the dichroism analysis.


CONCLUSIONS
We used circular dichroism and molecular modeling to examine the structure of ZNF706. The results suggest that human recombinant ZNF706 keeps its secondary structures and is appropriate for functional and structural studies. The method of expressing ZNF706 protein used in this study can be used to direct various functional and structural studies that will contribute to the understanding of its function as well as its relationship with other biological molecules and its putative role in carcinogenesis.
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Jucimara
J
Colombo
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Paola Jocelan Scarin
PJS
Provazzi
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Marilia Freitas
MF
Calmon
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Lilian Campos
LC
Pires
Institute of Chemistry of São Carlos, Department of Chemistry and Molecular Physics, University of São Paulo - USP, CEP: 13560-970, São Carlos /SP, Brazil.
Nathália Campos
NC
Rodrigues
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Paulo
P
Petl
Department of Physics, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Marcelo Andrés
MA
Fossey
Department of Physics, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Fátima Pereira
FP
de Souza
Institute of Chemistry of São Carlos, Department of Chemistry and Molecular Physics, University of São Paulo - USP, CEP: 13560-970, São Carlos /SP, Brazil.
Fernanda
F
Canduri
Department of Biology, São Paulo State University - UNESP, CEP: 15054-000, São José do Rio Preto /SP, Brazil.
Paula
P
Rahal