Mapping of factor XIII solvent accessibility as a function of activation state using chemical modification methods.
The transglutaminase Factor XIII (FXIII) catalyzes the formation of covalent cross-links between adjacent noncovalently associated fibrin chains in blood coagulation. The resulting covalently cross-linked hard clot is much more mechanically stable and resistant to proteolytic degradation. FXIII is activated by the serine protease thrombin in the presence of calcium ions. Protein modification experiments involving the labeling of cysteine and lysine side chains of the enzyme were performed before and after activation of the enzyme in an effort to gain further insight into structural changes occurring during the activation of FXIII. The experiments revealed differences in the labeling patterns of nonactivated and activated FXIII. These differences result from the exposure or sequestration of specific cysteine or lysine residues when the enzyme is activated, either physiologically with thrombin or nonproteolytically by exposure to calcium. Of note is the acetylation of Lys 73 and Lys 221 upon activation. Both of these residues lie within possible substrate recognition regions of FXIII. The active site Cys 314 is consistently alkylated in the activated enzyme, as is Cys 409, located near the dimer interface. Within the beta-barrel 2 domain of FXIII, Cys 695 becomes alkylated in activated FXIII. Within the same domain, an acetylated Lys (677 or 678), which is observed in the zymogen, cannot be found in the activated enzyme. The results provide a more extensive view of FXIII activation than has been previously available.
Turner
Brian T
BT
Department of Chemistry, University of Louisville, Louisville, Kentucky 40292, USA.
Sabo
T Michael
TM
Wilding
Diana
D
Maurer
Muriel C
MC
D000089:Acetic Anhydrides / Q000737:chemistry; D000107:Acetylation; D000478:Alkylation; D000595:Amino Acid Sequence; D003545:Cysteine / Q000737:chemistry; D005033:Ethylmaleimide / Q000737:chemistry; D005176:Factor XIII / Q000737:chemistry* / Q000378:metabolism; D026122:Factor XIIIa / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D036341:Intercellular Signaling Peptides and Proteins; D007460:Iodoacetamide / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D012996:Solutions; D012997:Solvents; D014357:Trypsin / Q000737:chemistry
D000089:Acetic Anhydrides; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D012996:Solutions; D012997:Solvents; C061895:factor XIII activation peptide; C064015:factor XIII subunit A; C031800:acetic anhydride; D005176:Factor XIII; D026122:Factor XIIIa; D014357:Trypsin; D008239:Lysine; D003545:Cysteine; D005033:Ethylmaleimide; D007460:Iodoacetamide