Dr. Kevin Siebenlist's Research
Hemostasis is a complex series of cell/cell, cell/protein, and protein/protein interactions designed to stem blood loss. Fibrinogen is a tridomainal disulfide linked plasma protein comprised of two symmetrical halves. Each half composed of three polypeptide chains termed Aα (red), Bβ (blue), and γ (green). Human fibrinogen can be separated into two major fractions by DEAE chromatography, fibrinogen 1 (‘peak 1 fibrinogen’) and fibrinogen 2 (‘peak 2 fibrinogen’). The two fibrinogens differ from each other with respect to their γ chain composition. Fibrinogen 1 contains two γA chains which are comprised of 411 amino acids. Heterodimeric fibrinogen 2 molecules contain one γA and one γ´ chain.
Illustration of a fibrinogen molecule
The variant γ´ chain is longer (427 residues), and has a more anionic, carboxyl terminal sequence than the γA chain beyond position 408. For many years the physiological function of the γ´ chain was unknown. Recently factor XIII has been shown to bind specifically to γ´ chains in fibrinogen 2 and thrombin has been shown to bind to the anionic γ´ extension of fibrin 2. After a cascading series of zymogen activations the plasma protein fibrinogen is converted to fibrin by the thrombin mediated removal of four peptides, two from the Aα chain and two from Bβ. Fibrin assembly commences with formation of double stranded twisting fibrils, subsequently, lateral fibril associations occur, resulting in thick fibers. Concomitant with converting fibrinogen to fibrin, thrombin converts protransglutaminase (factor XIII) to an active transglutaminase (factor XIIIa). Factor XIIIa introduces covalent cross-links between appropriately positioned γ chains to form γ dimers and between α chains forming α polymers.
Illustration of a two stranded fibrin fibril
The interactions between fibrin(ogen) and factor XIII are incompletely understood. To this end I am actively investigating how factor XIII is carried in the plasma by fibrinogen 2, the activation of factor XIII to factor XIIIa by thrombin, the movement of the active enzyme through the fibrin matrix, the effects of γ tetramers and γ trimers on the properties of the fibrin matrix, the intrinsic transglutaminase activity of the zymogen form of factor XIII, and the incorporation of α2-plasmin inhibitor into fibrin(ogen) the the factor XIII zymogen. An in vitro fibrinogen and factor XIII expression system is in the process of being established. Using these systems and site directed mutagenesis to introduce specific changes into the proteins the interactions between these molecules will be probed. Studies designed to elucidate the interactions between fibrin(ogen)/fibrin(ogen) and thrombin/fibrin will also use the in vitro fibrinogen expression system. These latter studies are being performed in collaboration with the Fibrinogen Research Laboratory (Dr. M.W. Mosesson) at the Blood Research Institute of The Blood Center of Southeastern Wisconsin.
- Meh DA, Mosesson MW, DiOrio JP, Siebenlist KR, Hernandez I, Amrani DL, and Stojanovich L. Disintegration and Reorganization of Fribrin Networks During TPA-Induced Clot Lysis, (2001) Blood Coag Fibrinol 12, 627-637.
- Mosesson MW, Siebenlist KR, Hernandez I, Wall JS, and Hainfeld JF. Fibrin Assembly and Crosslinking on a Fibrin Fragmen E Template, (2002) Thromb Haemostas 87, 651-658.
- Siebenlist KR. A Rebuttal: Cross-linking of Fibrinogen by Factor XIII Zymogen is not Apparent in Vivo, (2003) Thromb Haemostas 89, 944-945.
- Rosenthal AK, Mosesson MW, Gohr CM, Masuda I, Heinkel D, and Siebenlist KR. Regulation of Transglutaminase Activity in Articular Chondrocytes Through Thrombin Receptor-Mediated Factor XIII Synthesis, (2004) Thromb Haemostas 91, 558-568.
- Mosesson MW, Hernandez I, and Siebenlist KR. Evidence that Catalytically-Inactivated Thrombin Forms Dimers That Bridge Between Fibrin/Fibrinogen Fibers and Enhance Polymerization, (2004) Biophys Chem 110, 93-100.
- Siebenlist KR, Mosesson MW, Hernandez I, Bush LA, DiCera E, Shainoff JR, DiOrio JP, and Stojanovic L. Studies on the Basis for the Properties of Fibrin Produced From Fibrinogen-Containing Gamma' Chains, (2005) Blood 106, 2730-2736.
- Siebenlist KR. Response: Fibrinogen Containing y' Chains; Is the Assay Measuring What Farrell's Group Expects?, (2006) Blood 107, 3011-3012.
- Brennan SO, Mosesson MW, Lowen R, Siebenlist KR, Matsunaga A. Hypofibrinogenaemia Resulting from Novel Single Nucleotide Deletion at Condon Bbeta58 (3404del A) Associated with Thrombotic Stroke in Infancy, (2006) Thromb Haemost. 95, 738-739.
- Mosesson MW, Siebenlist KR, Hernandez I, Lee KN, Christiansen VJ, McKee PA. Evidence That α2-Antiplasmin Becomes Covalently Ligated to Plasma Fibrinogen in the Circulation: A New Role for Plasma Factor XIII in Fibrinolysis Regulation, (2008) J Thromb Haemost 6, 1565-1570.
- Siebenlist KR, Behm RA, Mosesson MW, Ariens R. Differential Cross-Linking Activity of the VAL34 and LEU34 Factor XIII Variants (In Preparation).