Purification, identification, and characterization of elastase on erythrocyte membrane as factor IX-activating enzyme

• Published in: Biochemical and biophysical research communications 2004-03, Vol.316 (1), p.65-70
• Main Authors: Iwata, Hiroki, Kaibara, Makoto, Dohmae, Naoshi, Takio, Koji, Himeno, Ryutaro, Kawakami, Satoshi
• Format: Article
• Language: English
• Subjects: Adult; Amino Acid Sequence; Blood Coagulation; Coagulation; Elastase; Erythrocyte; Erythrocyte Membrane - enzymology; Factor IX - chemistry; Factor IX - metabolism; Factor IX activation; Humans; Male; Molecular Sequence Data; Pancreatic Elastase - chemistry; Pancreatic Elastase - isolation & purification; Pancreatic Elastase - metabolism
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2004.02.020

In our previous papers, we reported that factor IX (F-IX), when activated by erythrocyte membranes, causes coagulation. We report on purification, identification, and characterization of F-IX-activating enzyme extracted from human erythrocyte membranes. The enzyme whose amino acid sequence is almost in accord with neutrophil elastase was found in normal erythrocyte membrane. The molecular mass was slightly smaller than that of neutrophil elastase. The content of the enzyme in erythrocyte membranes was estimated to be 3.0–3.7 ng per 10 6 erythrocytes. The F-IX sites cleaved by the enzyme were slightly different from those by the ordinary coagulation reaction. The ability of F-IX cleaved by the enzyme to cause coagulation was estimated to be approximately 1/10 as high as that of the F-IX cleaved by activated F-XI. These findings provide evidence that F-IX is activated by erythrocyte membrane, which may serve as a triggering mechanism for blood coagulation.