A new fibrinolytic enzyme (55 kDa) from Allium tuberosum: purification, characterization, and comparison.
Chives have been used both as food and as medicine. Previously, two fibrinolytic enzymes, ATFE-I (90 kDa) and ATFE-II (55 kDa), were identified in chives (Allium tuberosum), a perennial herb. In the present work, ATFE-II was purified by ion-exchange chromatography followed by gel filtration. In addition, the enzyme properties of ATFE-I and ATFE-II were compared. The molecular mass and isoelectric point (pI value) of ATFE-II were 55 kDa and pI 4.0, respectively, as revealed using one- or two-dimensional fibrin zymography. ATFE-II was optimally active at pH 7.0 and 45°C. ATFE-II degraded the Aα-chain of human fibrinogen but did not hydrolyze the Bβ-chain or the γ-chain, indicating that the enzyme is an α-fibrinogenase. The proteolytic activity of ATFE-II was completely inhibited by 1 mM leupeptin, indicating that the enzyme belongs to the cysteine protease class. ATFE-II was also inhibited by 1 mM Fe²(+). ATFE-II exhibited high specificity for MeO-Suc-Arg-Pro-Tyr-p-nitroaniline (S-2586), a synthetic chromogenic substrate of chymotrypsin. Thus proteolytic enzymes from A. tuberosum may be useful as thrombolytic agents.
Chung
Dong-Min
DM
Microbe-Based Fusion Technology Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk, Republic of Korea.
Choi
Nack-Shick
NS
Chun
Hyo Kon
HK
Maeng
Pil Jae
PJ
Park
Sang-Bong
SB
Kim
Sung-Ho
SH
D027782:Chive / Q000201:enzymology*; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D055808:Drug Discovery; D005296:Ferrous Compounds / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism; D005342:Fibrinolysis; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007526:Isoelectric Point; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D007976:Leupeptins / Q000494:pharmacology; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D035261:Plant Components, Aerial / Q000201:enzymology*; D010940:Plant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D015912:Thrombolytic Therapy; D013927:Thrombosis / Q000188:drug therapy
D015853:Cysteine Proteinase Inhibitors; D005296:Ferrous Compounds; D005343:Fibrinolytic Agents; D007527:Isoenzymes; D007976:Leupeptins; D009842:Oligopeptides; D010940:Plant Proteins; C464794:fibrinogen Aalpha; D005340:Fibrinogen; D003546:Cysteine Endopeptidases; C029451:ferrous chloride