Characterization of recombinant factor XIIIa produced in Saccharomyces cerevisiae.
Recombinant factor XIIIa (FXIIIa), produced in Saccharomyces cerevisiae, was recovered as a fully active cytosolic component and rigorously compared to natural F XIIIa from human placenta with respect to physicochemical and functional properties. Identical parameters were found in SDS polyacrylamide gel electrophoresis, analytical ultracentrifugation and HPLC gel filtration, and all spectral characteristics including derivative UV absorbance, fluorescence and circular dichroism were identical. Similarly, the interaction of both proteins with polyclonal antibodies directed against the entire FXIIIa or its N-terminal 4 kD activation peptide were identical. Furthermore, thrombin cleavage and fibrin cross-linking showed indistinguishable patterns. The only difference we observed was with respect to endgroup analysis. The recombinant protein is homogeneous, whereas placental FXIIIa shows multiple electrophoretic bands caused by microheterogeneity in the C-terminal part of the protein.
Rinas
U
U
Institut für Biophysik und Physikalische Biochemie, Universität Regensburg, FRG.
Risse
B
B
Jaenicke
R
R
Bröker
M
M
Karges
H E
HE
Küpper
H A
HA
Zettlmeissl
G
G
D003001:Cloning, Molecular; D003432:Cross-Linking Reagents / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D008970:Molecular Weight; D010920:Placenta / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013056:Spectrophotometry, Ultraviolet; D013917:Thrombin / Q000378:metabolism; D011503:Transglutaminases / Q000096:biosynthesis / Q000235:genetics*
D003432:Cross-Linking Reagents; D011994:Recombinant Proteins; D005337:Fibrin; D011503:Transglutaminases; D013917:Thrombin