Generation of monoclonal antibodies specific of the postfusion conformation of the Pneumovirinae fusion (F) protein.
Paramyxovirus entry into cells requires fusion of the viral and cell membranes mediated by one of the major virus glycoproteins, the fusion (F) glycoprotein which transits from a metastable pre-fusion conformation to a highly stable post-fusion structure during the membrane fusion process. F protein refolding involves large conformational changes of the protein trimer. One of these changes results in assembly of two heptad repeat sequences (HRA and HRB) from each protomer into a six-helix bundle (6HB) motif. To assist in distinguishing pre- and post-fusion conformations of the Pneumovirinae F proteins, and as extension of previous work (Palomo et al., 2014), a general strategy was designed to obtain polyclonal and particularly monoclonal antibodies specific of the 6HB motif of the Pneumovirinae fusion protein. The antibodies reported here should assist in the characterization of the structural changes that the F protein of human metapneumovirus or respiratory syncytial virus experiences during the process of membrane fusion.
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain.
Laura
L
Rodríguez
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain.
Eduardo
E
Olmedillas
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain.
Vicente
V
Mas
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain.
Mónica
M
Vázquez
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain.
Olga
O
Cano
Unidad de Microscopía Electrónica y Confocal, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
María C
MC
Terrón
Unidad de Microscopía Electrónica y Confocal, Instituto de Salud Carlos III, Majadahonda, 28220 Madrid, Spain.
Daniel
D
Luque
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain. Electronic address: cpalomo@isciii.es.
Concepción
C
Palomo
Unidad de Biología Viral, Centro Nacional de Microbiología and CIBER de Enfermedades Respiratorias, 28220 Madrid, Spain. Electronic address: jmelero@isciii.es.
José A
JA
Melero
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000302:isolation & purification*; D005260:Female; D008807:Mice, Inbred BALB C; D018099:Pneumovirinae / Q000276:immunology*; D011487:Protein Conformation; D011817:Rabbits; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014760:Viral Fusion Proteins
Conformation specific antibodies; Fusion proteins; Pneumovirinae