Literature summary for 3.4.21.68 extracted from

Liu, H.; Zhou, X.; Zhang, Y.
A comparative investigation on different refolding strategies of recombinant human tissue-type plasminogen activator derivative (2006), Biotechnol. Lett., 28, 457-463.

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Activating Compound

Activating Compound	Comment	Organism	Structure
cyanogen bromide-treated fibrinogen (CNBr-Fbg)	CNBr-Fbg (50 microgram/ml) give a 10-fold enhancement of activation, and addition of 2.86 microg/ml oversulfated chondroitin-6-sulfate or oversulfated fucoidan amplified this to 15-fold. A 25-fold to 35-fold enhancement of activation of glutamic plasminogen by tPA is obtained when 2.86 microgram/ml oversulfated compounds are combined with 16.2 mmol/l lysine and 50 microgram/ml CNBr-Fbg	Homo sapiens
L-lysine	addition of 16.2 mmol/l L-lysine give 3fold to 4fold enhancement of activation, which is further enhanced to 5fold to 6fold by addition of 2.86 microgram/ml oversulfated chondroitin-6-sulfate or oversulfated fucoidan	Homo sapiens
oversulfated chondroitin-6-sulfate	addition of 28.6 microgram/ml gives 2-fold to 4-fold increase in the rate of enhancement of activation of glutamic plasminogen by tPA using 0.05 mol/l Tris buffer (pH 7.35) containing NaCl (0.9%)	Homo sapiens
oversulfated fucoidan	addition of 28.6 microgram/ml gives 2-fold to 4-fold increase in the rate of enhancement of activation of glutamic plasminogen by tPA using 0.05 mol/l Tris buffer (pH 7.35) containing NaCl (0.9%)	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
additional information	recombinant human tissue-type plasminogen activator derivative (r-PA) (deletion mutant lacking the 4th-175th amino acid residues of t-PA), fused with thioredoxin (Trx), is expressed in Escherichia coli. Trx-r-PA fusion protein is almost completely expressed in the form of inclusion bodies and without activity. An optimized on-column refolding process for Trx-r-PA inclusion bodies is established: The collected Trx-r-PA inclusion bodies are dissolved in 6 M guanidine hydrochloride, and the denatured protein is separated from dithiothreitol and guanidine hydrochloride with a G25 column and simultaneously dissolved in 8 M urea containing oxidized glutathione. Refolding of Trx-r-PA protein on Sephacryl S-200 column with a decreasing urea gradient combined with two-stage temperature control is employed, and the activity recovery of refolded protein is increased from 3.6 to 13.8% in comparison with the usual dilution refolding	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Glu-Phe-Lys-p-nitroanilide + H2O	-	Homo sapiens	D-Glu-Phe-Lys + p-nitroaniline	-	?

Synonyms

Synonyms	Comment	Organism
human tissue-type plasminogen activator	-	Homo sapiens
tPA	-	Homo sapiens

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Release 2023.1 (January 2023)