Abstract
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.
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References
Jaenicke, R. and Rudolph, R. 1986. Folding and association of oligomeric proteins. Meth. Enzymol. 131: 218–250.
Schmid, F.X. 1989. Spectral methods of characterizing protein conformation and conformational changes, p. 251–285 In: Protein Structure. A Practical Approach, Vol. 1. Creighton, T. E. (Ed.). IRL Press, Oxford.
Delvos, U. and Müller-Berghaus, G. 1985. Die regulation der Blutgerinnung. Naturwissenschaften 72: 461–469.
Folk, J.E. and Finlayson, J.S. 1977. The (γ-glutamyl) lysin cross-link and the catalytic role of transaminases. Adv. Protein Chem. 31: 1–133.
Bohn, H. 1978. The human fibrin-stabilizing factors. Mol. Cell. Biochem. 20: 67–75.
Schwartz, M.L., Pizzo, S.V., Hill, R.L. and McKee, P.A. 1973. Human factor XIII from plasma and platelets. J. Biol. Chem. 248: 1395–1407.
Takahashi, N., Takahashi, Y. and Putnam, F.W. 1986. Primary structure of blood coagulation factor XHIa (fibrinoligase transglutaminase) from human placenta. Proc. Natl. Acad. Sci. USA 83: 8021–8023.
Ichinose, A., Hendrickson, L.E., Fujikawa, K. and Davie, E.W. 1986. Amino acid sequence of the a-subunit of human factor XIII. Biochemistry 25: 6900–6906.
Grundmann, U., Amann, E., Zettlmeissl, G. and Küpper, H.A. 1986. Characterization of cDNA coding for human factor XIIIa. Proc. Natl. Acad. Sci. USA 83: 8024–8028.
Amann, E., Abel, K.-J., Grundmann, U., Okazaki, H. and Küpper, H.A. 1988. Synthesis of human FXIIIa in bacterial cells. Behring Inst. Mitt. 82: 35–42.
Rinas, U., Risse, B., Jaenicke, R., Abel, K.-J. and Zettlmeissl, G. 1990. Denaturation-renaturation of FXIIIa isolated from human placenta. Biological Chemistry, Hoppe Seyler 371: 49–56.
Bröker, M. and Karges, H.E. 1988. Expression of human factor XIIIa in yeast. Yeast 4: S142.
Ichinose, A. and Davie, E.W. 1988. Characterization of the gene for the a-subunit of human factor XIII, a blood coagulation factor. Proc. Natl. Acad. Sci. USA 85: 5829–5833.
Suzuki, K., Matsui, K., Ito, S., Fujita, K. and Matsumoto, H. 1988. Polymorphism of the a-subunit of coagulation factor XIII: Evidence for subtypes of the FXIIIA*1 and FXIIIA*2 alleles. Am. J. Hum. Genet. 43: 170–174.
Takagi, T. and Doolittle, R.F. 1974. Amino acid sequence studies on factor XIII and the peptide released during its activation by thrombin. Biochemistry 13: 750–756.
Arfin, S.M. and Bradshaw, R.A. 1988. Cotranslational processing and protein turnover in eukaryotic cells. Biochemistry 27: 7981–7984.
Rudolph, R. and Jaenicke, R. 1976. Kinetics of reassociation and reactivation of pig muscle lactate dehydrogenase after acid denaturation. Eur. J. Biochem. 63: 409–417.
Jaenicke, R. 1987. Folding and association of proteins. Progr. Biophys. Mol. Biol. 47: 117–237.
Bohn, H. and Schwick, H.-G. 1971. Isolierung und Charakterisierung eines fibrin-stabilisierenden Faktors aus menschlichen Plazenten. Arzneimittel Forsch. 21: 1432–1439.
Bohn, H. 1972. Comparative studies on the fibrin-stabilizing factors from human plasma, platelets and placentas. Ann. N.Y. Acad. Sci. 202: 256–272.
Cesarini, G. and Murray, J.A. 1987. Plasmid vectors carrying the replication origin of filamentous single-stranded phages, p. 135–154. In: Genetic Engineering, Vol. 9. Setlow, J. K. (Ed.). Plenum Press, New York.
Bröker, M. 1986. Vectors for regulated high-level expression of proteins fused to truncated forms of Escherichia coli β-galactosidase. Gene Anal. Techn. 3: 53–57.
Bröker, M. and Amann, E. 1986. pUC12-STOP: An expression vector with portable translation stop signals. Appl. Microbiol. Biotechnol. 23: 294–296.
Wetlaufer, D.B. 1962. Ultraviolet spectra of proteins and amino acids. Adv. Protein Chem. 17: 304–390.
Yphantis, D.A. 1964. Equilibrium ultracentrifugation of dilute solutions. Biochemistry 3: 297–317.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Margolis, J. and Wrigley, C.W. 1975. Improvement of pore gradient electrophoresis by increasing the degree of cross-linkage at high acrylamide concentrations. J. Chromatography 106: 204–209.
Tsang, V.C.W., Peralta, J.M. and Simons, A.R. 1983. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specification of antigens and antibodies separated by gel-electrophoresis. Meth. Enzymol. 92: 377–391.
Hayashi, R., 1976. Carboxypeptidase Y. Meth. Enzymol. 45: 568–587.
Qureshi, A.G., Fohlin, L. and Bergström, J. 1984. Application of high-performance liquid chromatography to the determination of free amino acids in physiological fluids. J. Chromatography 297: 91–100.
Egbring, R., Schmidt, W. and Havemann, K. 1973. Die vereinfachte radiologische Faktor XIII-Bestimmung und ihre klinische Anwendung bei kongenitalem FXIII-Mangel. Blut 27: 6–19.
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Rinas, U., Risse, B., Jaenicke, R. et al. Characterization of Recombinant Factor XIIIa Produced in Saccharomyces cerevisiae. Nat Biotechnol 8, 543–546 (1990). https://doi.org/10.1038/nbt0690-543
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DOI: https://doi.org/10.1038/nbt0690-543