Novel Aalpha chain truncation (fibrinogen Perth) resulting in low expression and impaired fibrinogen polymerization.
A young woman with a history of menorrhagia and easy bruising presented with a functional fibrinogen concentration of 1.8 mg mL(-1), a gravimetric concentration of 3.3 mg mL(-1) and a prolonged thrombin clotting time of 32 s. Both reverse phase analysis and reducing SDS-PAGE revealed a normal profile of Aalpha, Bbeta, and gamma chains. However, non-reducing gels revealed a broadened 340-kDa band, while the 305-kDa band was normal, suggesting a C-terminal truncation of the Aalpha chain. DNA sequencing of all exons and intron boundaries revealed a single heterozygous cytosine deletion at nucleotide 4841 of the Aalpha gene predicting a frameshift and the incorporation of 23 new residues (LMKLPSSTLPQLEKHSQVSSHLC) before termination after residue 517. In agreement with a predicted mass decrease of 9953 Da, the measured mass of the Aalpha(Perth) chain was 56 242 Da, while that of the normal Aalpha(A) chain was 66 189 Da. Tryptic mapping of isolated Aalpha chains revealed a new [M + 2H] ion at 607 m z(-1), corresponding to the predicted penultimate peptide LPSSTLPQLEK. The variant chain was poorly incorporated into plasma fibrinogen at a ratio of Aalpha(Perth)/Aalpha(A) of 0.15 : 1, suggesting the Aalpha(Perth) chain might be out-competed by normal chains during molecular assembly in the hepatocyte. Despite the low expression, polymerization curves showed a decreased V(max) and final turbidity, suggesting the fibrinogen Perth clots are composed of thinner fibers. However, the fibrinolytic rate was very similar to that of the control.
Homer
V M
VM
Molecular Pathology Laboratory, Canterbury Health Laboratories, Christchurch Hospital, Christchurch, New Zealand. vivienne.homer@chmeds.ac.nz
Mullin
J L
JL
Brennan
S O
SO
Barr
A
A
George
P M
PM
D000328:Adult; D000595:Amino Acid Sequence; D001777:Blood Coagulation / Q000235:genetics; D004252:DNA Mutational Analysis; D005192:Family Health; D005260:Female; D015241:Fibrinogens, Abnormal / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D005342:Fibrinolysis / Q000235:genetics; D016368:Frameshift Mutation; D006579:Heterozygote; D006801:Humans; D007700:Kinetics; D008595:Menorrhagia / Q000235:genetics; D017384:Sequence Deletion
D015241:Fibrinogens, Abnormal; C483329:fibrinogen Perth