A novel fibrinolytic enzyme extracted from the earthworm, Lumbricus rubellus.
A strong fibrinolytic enzyme was readily obtained in saline extracts of the earthworm, Lumbricus rubellus. It hydrolyzed not only plasminogen-rich fibrin plates, but also plasminogen-free fibrin plates. The average fibrinolytic activity was about 100 CU (plasmin units) or 250 IU (urokinase units)/g wet weight. The molecular weight and isoelectric point were about 20,000 and 3.4, respectively. The enzyme was heat-stable and displayed a very broad optimal pH range. DFP and SBTI strongly inhibited the enzyme, but the anti-plasmin agent, t-AMCHA, exerted little effect under the same conditions. Purification of the enzyme was performed and three partially purified fractions were obtained. These three fractions were further subdivided. The first fraction (F-I) was divided into three fractions (F-I-0, F-I-1, and F-I-2), which exhibited similar biochemical characteristics. The second fraction (F-II) could not be subdivided. The third fraction (F-III) was divided into two fractions (F-III-1 and F-III-2). Based on results for their enzymatic activities against various substrates, the fraction I enzymes are thought to represent a chymotrypsin-like enzyme and the fraction III enzymes to represent a trypsin-like enzyme. The fraction II enzyme appears to be neither a trypsin- or chymotrypsin-like enzyme nor an elastase. The amino acid compositions of the six enzymes were estimated. Compared with other serine enzymes, these enzymes contained very abundant asparagine or aspartic acid, and there was very little proline or lysine. From the above data, these enzymes are regarded as novel fibrinolytic enzymes, and we name them collectively as Lumbrokinase from the generic name of the earthworm.
Mihara
H
H
Department of Physiology, Miyazaki Medical College, Japan.
Sumi
H
H
Yoneta
T
T
Mizumoto
H
H
Ikeda
R
R
Seiki
M
M
Maruyama
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000933:Antifibrinolytic Agents / Q000494:pharmacology; D005342:Fibrinolysis; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009835:Oligochaeta / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature; D013997:Time Factors; D014020:Tissue Extracts / Q000737:chemistry / Q000494:pharmacology / Q000502:physiology*; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology
D000596:Amino Acids; D000933:Antifibrinolytic Agents; D014020:Tissue Extracts; D010447:Peptide Hydrolases; D014568:Urokinase-Type Plasminogen Activator