Sushi domains in the B subunit of factor XIII responsible for oligomer assembly.
Factor XIII (FXIII) is a heterotetramer composed of two catalytic A subunits (FXIII-A) and two B subunits (FXIII-B). FXIII-B has 10 Sushi domains. To explore the structure-function relationship of FXIII-B, we looked for domains in FXIII-B responsible for its homodimer and heterotetramer assembly with FXIII-A. Full-length recombinant human FXIII-B (rFXIII-B) and truncated rFXIII-Bs with various numbers of Sushi domains (rFXIII-B x- y ) were expressed in a baculovirus expression system. rFXIII-B was indistinguishable from purified human plasma FXIII-B, in terms of the molecular weight (after being deglycosylated by glycosidases) and the ability to form complexes between the two subunits. rFXIII-B was in dimer form and produced a heterotetramer complex with FXIII-A. Gel-filtration and FXIII-A binding analysis of the various truncated forms of rFXIII-B x- y revealed that the first Sushi domain was responsible for the binding of FXIII-B to FXIII-A and that the fourth and ninth Sushi domains were involved in the FXIII-B homodimer assembly. rFXIII-B and rFXIII-B 1-9, which formed a heterotetramer complex with FXIII-A, protected FXIII-A from proteolytic digestion. These findings suggest that only full-length or nearly full-length FXIII-B is large enough to cover the exposed surface of FXIII-A. In conclusion, at least 3 out of the 10 Sushi domains of FXIII-B have the distinct function of forming a homodimer and a heterotetramer, which should be ascribed to the differences in their amino acid sequences. The present studies, however, do not exclude the possibility that additional Sushi domains may also support either or both functions.
Souri
Masayoshi
M
Department of Molecular Patho-Biochemistry and Patho-Biology on Blood and Circulation, Yamagata University School of Medicine, Yamagata, Japan.
Kaetsu
Hiroshi
H
Ichinose
Akitada
A
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D005176:Factor XIII / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins; D018411:Spodoptera / Q000166:cytology
D011994:Recombinant Proteins; C064015:factor XIII subunit A; D005176:Factor XIII; C054380:factor XIIIb