Abstract
The strongest fibrinolytic protease (F-III-2) in the six enzyme proteins purified from earthworm, Lumbricus rubellus [N. Nakajima et al., Biosci. Biotech. Biochem., 57, 1726-1730 (1993)] has been modified chemically with fragmented human serum albumin (mol. wt., 10,000-30,000). The modified enzyme lost the antigenicity of the native enzyme and reacted with the antisera against human serum albumin, the human serum albumin fragments, and the conjugate with the native enzyme to form precipitation lines, which fused with each other. The conjugate was significantly more resistant to inactivation by protease inhibitors in rat plasma. The enzyme was a non-hemorrhagic protein and did not induce platelet aggregation. The enzyme kept potent proteolytic activity for fibrin and fibrinogen than that of human plasmin. The enzyme easily solubilized actual fibrin clots (thrombi) of whole blood induced by thrombin in a rat's vena cava. The continuous fibrinolysis for fibrin suspension in an enzyme reactor system using the modified enzyme immobilized to oxirane-activated acrylic beads has been achieved without any inactivation of the activity at least for more than 1 month. The N-terminal amino acid sequence of the protein was also investigated and the sequence showed local similarity to those of the serine proteases such as plasmin and chymotrypsin.
| Original language | English |
|---|---|
| Pages (from-to) | 293-300 |
| Number of pages | 8 |
| Journal | Bioscience, Biotechnology and Biochemistry |
| Volume | 60 |
| Issue number | 2 |
| Publication status | Published - Feb 1996 |
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Keywords
- Chemical modification
- Fibrinolytic enzyme (of earthworm)
- Immobilized enzyme
- Protein sequence
- Serine protease (of earthworm)
ASJC Scopus subject areas
- Food Science
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biotechnology
- Chemistry (miscellaneous)
- Applied Microbiology and Biotechnology
Cite this
Chemical Modification of Earthworm Fibrinolytic Enzyme with Human Serum Albumin Fragment and Characterization of the Protease as a Therapeutic Enzyme. / Nakajima, Nobuyoshi; Ishihara, Kohji; Sugimoto, Manabu; Sumi, Hiroyuki; Mikuni, Katsuhiko; Hamada, Hiroki.
In: Bioscience, Biotechnology and Biochemistry, Vol. 60, No. 2, 02.1996, p. 293-300.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Chemical Modification of Earthworm Fibrinolytic Enzyme with Human Serum Albumin Fragment and Characterization of the Protease as a Therapeutic Enzyme
AU - Nakajima, Nobuyoshi
AU - Ishihara, Kohji
AU - Sugimoto, Manabu
AU - Sumi, Hiroyuki
AU - Mikuni, Katsuhiko
AU - Hamada, Hiroki
PY - 1996/2
Y1 - 1996/2
N2 - The strongest fibrinolytic protease (F-III-2) in the six enzyme proteins purified from earthworm, Lumbricus rubellus [N. Nakajima et al., Biosci. Biotech. Biochem., 57, 1726-1730 (1993)] has been modified chemically with fragmented human serum albumin (mol. wt., 10,000-30,000). The modified enzyme lost the antigenicity of the native enzyme and reacted with the antisera against human serum albumin, the human serum albumin fragments, and the conjugate with the native enzyme to form precipitation lines, which fused with each other. The conjugate was significantly more resistant to inactivation by protease inhibitors in rat plasma. The enzyme was a non-hemorrhagic protein and did not induce platelet aggregation. The enzyme kept potent proteolytic activity for fibrin and fibrinogen than that of human plasmin. The enzyme easily solubilized actual fibrin clots (thrombi) of whole blood induced by thrombin in a rat's vena cava. The continuous fibrinolysis for fibrin suspension in an enzyme reactor system using the modified enzyme immobilized to oxirane-activated acrylic beads has been achieved without any inactivation of the activity at least for more than 1 month. The N-terminal amino acid sequence of the protein was also investigated and the sequence showed local similarity to those of the serine proteases such as plasmin and chymotrypsin.
AB - The strongest fibrinolytic protease (F-III-2) in the six enzyme proteins purified from earthworm, Lumbricus rubellus [N. Nakajima et al., Biosci. Biotech. Biochem., 57, 1726-1730 (1993)] has been modified chemically with fragmented human serum albumin (mol. wt., 10,000-30,000). The modified enzyme lost the antigenicity of the native enzyme and reacted with the antisera against human serum albumin, the human serum albumin fragments, and the conjugate with the native enzyme to form precipitation lines, which fused with each other. The conjugate was significantly more resistant to inactivation by protease inhibitors in rat plasma. The enzyme was a non-hemorrhagic protein and did not induce platelet aggregation. The enzyme kept potent proteolytic activity for fibrin and fibrinogen than that of human plasmin. The enzyme easily solubilized actual fibrin clots (thrombi) of whole blood induced by thrombin in a rat's vena cava. The continuous fibrinolysis for fibrin suspension in an enzyme reactor system using the modified enzyme immobilized to oxirane-activated acrylic beads has been achieved without any inactivation of the activity at least for more than 1 month. The N-terminal amino acid sequence of the protein was also investigated and the sequence showed local similarity to those of the serine proteases such as plasmin and chymotrypsin.
KW - Chemical modification
KW - Fibrinolytic enzyme (of earthworm)
KW - Immobilized enzyme
KW - Protein sequence
KW - Serine protease (of earthworm)
UR - http://www.scopus.com/inward/record.url?scp=0030087823&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030087823&partnerID=8YFLogxK
M3 - Article
C2 - 9063978
AN - SCOPUS:0030087823
VL - 60
SP - 293
EP - 300
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 2
ER -