Plasmin

Plasminogen:
Plasminogen is the precursor to the fibronolytic protease plasmin as well as angiostatin, an angiogenesis/tumor metastasis inhibitor. Human prostate carcinoma cells release urokinase and free sulfhydryl donors which can produce angiostatin from plasminogen. In vitro, urokinase, tissue plasminogen activator, or streptokinase, in combination with sulfydryl donors, such as N-acetyl-L-cysteine, D-penicillamine, captopril, L-cysteine, or reduced glutathione, produced angiostatin from plasminogen.1
Native-intact human plasminogen is a 291 amino acid glyco-protein with as many as 24 disulfide bonds. Plasminogen contains a single N-linked sialylated biantennary glycan. The two O-glycans possess a Gal β-1-3GalNAc core which is α-2-3 sialylated at the terminal Gal. An additional disialylated form has a second sialic acid residue with an α-2-6 linkage to GalNAc. Mono- and disialylated forms occur at a molar ratio of 80:20 in human plasminogen.2,3
Plasmin:
MW: ~86 kDa4 (Human)
When run on SDS-PAGE, bands appeared at 68, 46.2, 23.1 and 12.3 kDa.
Plasmin is a serine endopeptidase of the peptidase S1 family. Plasmin is converted to plasminogen by cleavage between Arg561 and Val562. The resulting activated plasmin consists of two disulfide-linked polypeptide chains. The plasmin heavy chain (MW 60 kDa) is derived from the amino terminal region of plasminogen. The light chain originates from the carboxyl-terminus of plasminogen. In vivo, the MW of the heavy chain can vary from 63KDa to 12KDa depending on the extent of proteolysis to the plasminogen from which it is derived. Plasmin is activated by a variety of proteases including urokinase, tissue plasminogen activator, and streptokinase. During the activation of plasminogen, an autolytic peptide of molecular weight 8,200 is released from the Glu-amino terminus to yield a Lys-amino terminus on the heavy chain. The active site of plasmin is on the light chain.1
Specificity and Kinetics
Plasmin exhibits preferential cleavage at the carboxyl side of Lysine and Arginine residues with higher selectivity than trypsin.5 It converts polymerized fibrin into soluble products.

pH Optimum: 8.5
pH 7.5: about 40% of maximal activity, pH 9.5: about 50% of maximal activity6

Temperature Optimum: 37 °C with rapid inactivation at 56 °C6
Plasmin is readily soluble in water (1 mg/ml). Solutions should be kept on ice and discarded at the end of the day or aliquoted and frozen at a concentration of 1 mg/ml. Frozen solutions will lose approximately 10% activity per month.
  • Gatley, S., et al., Proc. Natl. Acad. Sci. U S A, 94, 10868–10872 (1997)
  • Marti, T., et al., Eur J Biochem., 173, 57-63 (1988)
  • Pirie-Shepherd, S.R., et al., J Biol. Chem.,272, 7408-11 (1997)
  • Trends Biochem. Sci., 4, 1 (1979)
  • IUBMB Enzyme Nomenclature: www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/7.html
  • B.H.W., et al., Biochim. Biophys. Acta, 67, 658-668 (1963)
Plasminogen in vivo Processing
Lys-Plasmin

Plasmin Inhibitors

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A2504 6-Aminocaproic acid ≥99% (titration), powder
A7824 6-Aminocaproic acid BioUltra, ≥99%
A8456 4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride ≥97%
A6191 Antipain dihydrochloride from microbial source protease inhibitor
A8849 α2-Antiplasmin from human plasma lyophilized powder, ≥5 units/mg protein
A9141 Antithrombin III from bovine plasma lyophilized powder, 200-400 units/mg protein (E1%/280 = 6.5)
A2221 Antithrombin III from human plasma lyophilized powder, ≥95% (SDS-PAGE)
A1153 Aprotinin from bovine lung lyophilized powder, 3-8 TIU/mg solid
E0518 C1 Esterase Inhibitor from human plasma aqueous solution, ≥95% (SDS-PAGE)
D7910 3,4-Dichloroisocoumarin serine protease inhibitor
D0879 Diisopropylfluorophosphate
E0881 Elastatinal microbial
G2417 Gabexate mesylate analytical standard, for drug analysis
L2023 Leupeptin trifluoroacetate salt ≥90% (HPLC), microbial
M6159 α2-Macroglobulin from human plasma BioUltra, ≥98% (SDS-PAGE)
P7626 Phenylmethanesulfonyl fluoride ≥98.5% (GC)

Plasmin Substrates

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A8171 D-Ala-Leu-Lys-7-amido-4-methylcoumarin ≥95% (HPLC)
G8148 Gly-Arg-p-nitroanilide dihydrochloride unokinase and plasmin substrate
SCP0261 H-D-Ile-Phe-Lys-pNA trifluoroacetate salt New
N8010 4-Nitrophenyl 4-guanidinobenzoate hydrochloride protease inhibitor and substrate
T6140 N-(p-Tosyl)-Gly-Pro-Lys 4-nitroanilide acetate salt plasmin substrate
V0882 D-Val-Leu-Lys 4-nitroanilide dihydrochloride plasmin substrate

Products

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P1867 Plasmin from human plasma lyophilized powder, ≥2.0 units/mg protein
10602370001 Plasmin, bovine from bovine plasma
10602361001 Plasmin, human from human plasma
P9156 Plasminogen from bovine plasma lyophilized powder, ≥2.0 units/mg protein
P7397 Plasminogen from human plasma lyophilized powder, ≥2 units/mg protein

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F5386 Fibrin from human plasma insoluble powder
S3134 Streptokinase from β-hemolytic Streptococcus (Lancefield Group C) lyophilized powder, ≥3,000 units/mg solid
U0633 Urokinase from human urine lyophilized powder, ≥500 units/mg protein