Expression of functional human coagulation factor XIII A-domain in plant cell suspensions and whole plants.
Coagulation factor XIII, a zymogen present in blood as a tetramer (A2B2) of A- and B-domains, is one of the components of many "wound sealants" which are proposed for use or currently in use as effective hemostatic agents, sealants, and tissue adhesives in surgery. After activation by alpha-thrombin cleavage, coagulation factor XIII A-domain, a transglutaminase, is formed and catalyzes the covalent cross-linking of the alpha- and gamma-chains of linear fibrin to form homopolymers, which can quickly stop bleeding. We have successfully expressed the A-domain of factor XIII in both plant cell cultures and whole plants. Transgenic plant cell culture allows a rapid method for testing production feasibility while expression in whole plants demonstrates an economic production system for recombinant human plasma-based proteins. The expressed factor XIII A-domain had a similar size as that of human plasma-derived factor XIII. Crude plant extract containing recombinant factor XIII A-domain showed transglutaminase activity with monodansylcadaverine and casein as substrates and cross-linking activity in the presence of linear fibrin. The expression of factor XIII A-domain was not affected by plant leaf position.
Gao
Johnway
J
Battelle Memorial Institute, 902 Battelle Blvd., Richland, WA 99352, USA. gaoj@battelle.org
Hooker
Brian S
BS
Anderson
Daniel B
DB
D005176:Factor XIII / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005337:Fibrin / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008958:Models, Molecular; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology / Q000235:genetics / Q000502:physiology*
D011994:Recombinant Proteins; D005337:Fibrin; D005176:Factor XIII