Expression and purification of human respiratory syncytial virus recombinant fusion protein.
The Human Respiratory Syncytial Virus (HRSV) fusion protein (F) was expressed in Escherichia coli BL21A using the pET28a vector at 37 degrees C. The protein was purified from the soluble fraction using affinity resin. The structural quality of the recombinant fusion protein and the estimation of its secondary structure were obtained by circular dichroism. Structural models of the fusion protein presented 46% of the helices in agreement with the spectra by circular dichroism analysis. There are only few studies that succeeded in expressing the HRSV fusion protein in bacteria. This is a report on human fusion protein expression in E. coli and structure analysis, representing a step forward in the development of fusion protein F inhibitors and the production of antibodies.
Arcuri
Helen A
HA
Instituto de Biociências, Letras e Ciências Exatas, UNESP, Rua Cristóvão Colombo, São José do Rio Preto, SP, Brazil.
Apponi
Luciano H
LH
Valentini
Sandro R
SR
Durigon
Edison L
EL
de Azevedo
Walter F
WF
Fossey
Marcelo A
MA
Rahal
Paula
P
de Souza
Fatima P
FP
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D018113:Respiratory Syncytial Virus, Human / Q000737:chemistry*; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
C526850:F protein, human respiratory syncytial virus; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins