Porcine factor V: cDNA cloning, gene mapping, three-dimensional protein modeling of membrane binding sites and comparative anatomy of domains.
Factor V is a plasma protein essential for blood coagulation. This protein is involved in activated protein C resistance, the most common inherited thrombotic disorder known. We utilized the polymerase chain reaction to clone the porcine factor V gene by generating overlapping clones amplified with primers chosen by comparison with known nucleotide sequences. The porcine factor V cDNA contig encodes a predicted 2258-amino acid protein, making it the largest in comparison to the bovine, human, and murine proteins. Porcine factor V has the highest level of homology with bovine factor V, but also has high levels of conservation of important residues with all the species. Radiation hybrid mapping assigned the porcine factor V gene to chromosome 4. Three-dimensional models of factor V were generated and used to analyze membrane-binding sites in terms of conserved, and therefore likely important residues.
Shriners Hospital for Children, Center for Research in Skeletal Development and Pediatric Orthopedics, Special Shared Facility, Tampa, Florida 33612, USA. dgrimm@shctampa.usf.edu
D R
DR
Grimm
M B
MB
Colter
M
M
Braunschweig
L J
LJ
Alexander
P J
PJ
Neame
H K
HK
Kim
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers; D005165:Factor V / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021101:Radiation Hybrid Mapping; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000235:genetics*
D017931:DNA Primers; D005165:Factor V