Open AccessStructural and functional influences of coagulation factor XIII subunit B heterozygous missense mutants
Author(s) -
Thomas Anne,
Biswas Arijit,
Ivaskevicius Vytautas,
Oldenburg Johannes
Publication year - 2015
Publication title -
molecular genetics and genomic medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.765
H-Index - 29
ISSN - 2324-9269
DOI - 10.1002/mgg3.138
Subject(s) - heterotetramer , missense mutation , factor xiii , protein subunit , compound heterozygosity , chemistry , fibrin , phenotype , microbiology and biotechnology , biology , immunology , biochemistry , gene
Abstract The coagulation factor XIII ( FXIII ) is a plasma circulating heterotetrameric protransglutaminase that acts at the end of the coagulation cascade by covalently cross‐linking preformed fibrin clots (to themselves and to fibrinolytic inhibitors) in order to stabilize them against fibrinolysis. It circulates in the plasma as a heterotetramer composed of two homomeric catalytic Factor XIIIA 2 ( FXIIIA 2 ) and two homomeric protective/carrier Factor XIIIB 2 subunit ( FXIIIB 2 ). Congenital deficiency of FXIII is of two types: severe homozygous/compound heterozygous FXIII deficiency which results in severe bleeding symptoms and mild heterozygous FXIII deficiency which is associated with mild bleeding (only upon trauma) or an asymptomatic phenotype. Defects in the F13B gene (Factor XIIIB subunit) occur more frequently in mild FXIII deficiency patients than in severe FXIII deficiency. We had recently reported secretion‐related defects for seven previously reported F13B missense mutations. In the present study we further analyze the underlying molecular pathological mechanisms as well as the heterozygous expression phenotype for these mutations using a combination of in vitro heterologous expression (in HEK 293T cells) and confocal microscopy. In combination with the in vitro work we have also performed an in silico solvated molecular dynamic simulation study on previously reported FXIIIB subunit sushi domain homology models in order to predict the putative structure‐functional impact of these mutations. We were able to categorize the mutations into the following functional groups that: (1) affect antigenic stability as well as binding to FXIIIA subunit, that is, Cys5Arg , Cys316Phe , and Pro428Ser (2) affect binding to FXIIIA subunit with little or no influence on antigenic stability, that is, Ile81Asn and Val401Gln c) influence neither aspects and are most likely causality linked polymorphisms or functional polymorphisms, that is, Leu116Phe and Val217Ile . The Cys5Arg mutation was the only mutation to show a direct secretion‐based defect since the mutated protein was observed to accumulate in the endoplasmic reticulum.