caa a22 4500 397557833 CHVBK 20180308164808.0 cr unu---uuuuu 161202e199605 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021336 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021336 Expression and Release of the a and b Subunits for Human Coagulation Factor XIII in Baby Hamster Kidney (BHK) Cells [Elektronische Daten] [Hiroshi Kaetsu, Teruto Hashiguchi, Don Foster, kitada Ichinose] The a subunit of coagulation factor XIII lacks a hydrophobic signal sequence for secretion from cells, while the b subunit has a typical signal sequence. To determine whether the a subunit can be synthesized and released, expression vectors containing the cDNA for either subunit were transfected into baby hamster kidney (BHK) cells. Western blotting analysis and gel nitration chromatography demonstrated that the recombinant a and b subunits (rXUIa and rXIIIb) had the same molecular weights and subunit structures (a2, b2, and a2b2) as the native molecules. rXIIIa was enzymatically active when activated by thrombin. Most rXIIIb was secreted as measured by ELISA, while most rXIIIa was detected in the cytosol by subcellular fractionation. Co-expression with r XIIIb in the same cells did not promote the release of rXIIIa. Treatment of the cells with brefeldin A, a potent inhibitor of protein transportation, blocked the secretion of r XIIIb, although it had no effect on the release of rXIIIa. Several drugs and heat stress induced the release of rXIIIa, which correlated directly with that of cytoplasmic lactate dehydrogenase. These results suggest that the a subunit is released from cells as a consequence of cell injury, which is independent of the classical secretory pathway. © by the Journal of Biochemistry Regular Papers nationallicence biosynthesis nationallicence blood coagulation nationallicence factor XIII nationallicence secretory pathway nationallicence transglutaminase nationallicence Kaetsu Hiroshi Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut Hashiguchi Teruto Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut Foster Don ZymoGenetics, Inc Seattle, WA 98105, USA aut Ichinose kitada Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut The Journal of Biochemistry Oxford University Press 119/5(1996-05), 961-969 0021-924X 119:5<961 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021336 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021336 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kaetsu Hiroshi Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut NATIONALLICENCE 700 1- Hashiguchi Teruto Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut NATIONALLICENCE 700 1- Foster Don ZymoGenetics, Inc Seattle, WA 98105, USA aut NATIONALLICENCE 700 1- Ichinose kitada Department of Biochemistry, University of Washington Seattle, WA 98195, USA aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/5(1996-05), 961-969 0021-924X 119:5<961 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford