Cross-linking of protein subunits by 1,3, 5-triacryloyl-hexahydro-s-triazine.
Six difunctional and trifunctional derivatives of acrylamide were synthesized and investigated as potential protein lysine residue cross-linking agents. 1,3,5-Triacryloyl-hexahydro-s-triazine (TAT) was considered the best. The rate constants for the reactions of TAT with model nucleophiles in water solution at 25 degreesC were with the glycine anion amino group, 7.69 x 10(-3) M-1 s-1; with the anionic form of the N-acetyl-L-cysteine thiol group, 5.54 M-1 s-1; and with the Nalpha-acetyl-L-histidine imidazole ring, 1.19 x 10(-5) M-1 s-1 (at pH 9.0). The kinetics of modification of amino groups by TAT were studied for several proteins: alpha1-casein, bovine serum albumin, recombinant human growth hormone, recombinant human interferons-alpha2b, and -gamma. The results indicate that if proteins are associated into oligomeric structures in water, their subunits are effectively cross-linked by TAT.
Dienys
G
G
Faculty of Chemistry, Vilnius University, Lithuania, and Institute of Biotechnology, Graiciūno 8, Vilnius 2028, Lithuania.
Sereikaite
J
J
Gavenas
G
G
Kvederas
R
R
Bumelis
V A
VA
D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D006207:Half-Life; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D014227:Triazines; D014867:Water
D003432:Cross-Linking Reagents; D011506:Proteins; D012996:Solutions; D014227:Triazines; D014867:Water; C049485:1,3,5-triacryloylhexahydrotriazine