Specific adsorption of serine proteases on coated silica beads substituted with amidine derivatives.
Amidine derivatives interact with serine proteases, the inhibition being due to interactions between amidine functions and the active sites of the enzymes. Five different types of amidine (substituted or unsubstituted) were coupled to coated silica beads, which had previously been coated with DEAE-dextran to minimize the non-specific interactions due to silanol groups. Coated silica functionalized with substituted amidines shows a strong affinity towards human plasmin. This affinity is probably due to hydrophobic interactions between the substituted amidine and the human plasmin structure. Coated silica grafted by p-aminobenzamide gives a specific interaction with human plasmin. The importance of ionic strength and the steric conformation of the ligand is discussed. This support was used to purify thrombin from crude preparations by high-performance affinity chromatography.
Khamlichi
S
S
LRM, CNRS URA 502, Université Paris-Nord, Villetaneuse, France.
Muller
D
D
Fuks
R
R
Jozefonvicz
J
J
D000327:Adsorption; D000578:Amidines; D000818:Animals; D001120:Arginine; D001550:Benzamidines; D002417:Cattle; D002846:Chromatography, Affinity; D005341:Fibrinolysin / Q000302:isolation & purification; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D007202:Indicators and Reagents; D012697:Serine Endopeptidases / Q000302:isolation & purification*; D012822:Silicon Dioxide; D013816:Thermodynamics; D013917:Thrombin / Q000302:isolation & purification
D000578:Amidines; D001550:Benzamidines; D006146:Guanidines; D007202:Indicators and Reagents; C006500:4-aminobenzamidine; D012822:Silicon Dioxide; D001120:Arginine; D012697:Serine Endopeptidases; D013917:Thrombin; D005341:Fibrinolysin; D019791:Guanidine