Carbodiimide modification enhances activity of pig pancreatic phospholipase A2.

@article{Ferreira1994CarbodiimideME,
  title={Carbodiimide modification enhances activity of pig pancreatic phospholipase A2.},
  author={Jo{\~a}o Pinto Ferreira and Ram Sasisekharan and Omid Louie and Robert S. Langer},
  journal={European journal of biochemistry},
  year={1994},
  volume={223 2},
  pages={
          611-6
        }
}
Pig phospholipase A2, pig iso-phospholipase A2 and bovine pancreatic phospholipase A2 were reacted in solution with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, in the presence of N-hydroxysulfosuccinimide, at pH 7. The influence of micellar protectants was analyzed. In the presence of n-hexadecylphosphocholine, the losses of activity in micellar diheptanoyl-lecithin were 80, 35, and 10% in bovine phospholipase A2, pig iso-phospholipase A2, and pig phospholipase A2, respectively. With 1… 
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References

SHOWING 1-10 OF 19 REFERENCES
Modification of carboxylate groups in bovine pancreatic phospholipase A2. Identification of aspartate-49 as Ca2+-binding ligand.
TLDR
Carboxylase groups in bovine pancreatic phospholipase A2 were modified using a water-soluble carbodiimide and semicarbazide and the label was found to be attached to Asp-49, demonstrating the essential role of As p-49 in Ca2+ binding.
Influence of chemistry in immobilization of cobra venom phospholipase A2: implications as to mechanism.
TLDR
The results suggest that the enzyme activity lost upon immobilization is a result of the inherent chemical modification of the enzyme and that enzyme oligomerization and interfacial recognition are not cause-effect phenomena.
Glutamic acid 71 and aspartic acid 66 control the binding of the second calcium ion in porcine pancreatic phospholipase A2.
TLDR
The Gln92 mutant PLA displayed the same properties as native phospholipase indicating that Glu92 is not important for binding the second metal ion, however, Glu71 and, to a lesser extent, Asp66 are both directly involved in the low-affinity calcium binding.
Interfacial catalysis by phospholipase A2: monomeric enzyme is fully catalytically active at the bilayer interface.
TLDR
It was concluded that secretory PLA2s from venoms and pancreas are fully catalytically active as monomeric PLA2, and that acylation of PLA2 was not necessary for catalysis or binding to the interface and that the binding of the substrate to the active site of PLA1 was not needed.
Enhanced activity and altered specificity of phospholipase A2 by deletion of a surface loop.
TLDR
Removal of residues 62 to 66 from porcine pancreatic phospholipase A2 does not change the binding constant for micelles significantly, but it improves catalytic activity up to 16 times on micellar (zwitterionic) lecithin substrates.
Phospholipase A2 engineering. X-ray structural and functional evidence for the interaction of lysine-56 with substrates.
TLDR
The results suggest that the side chain of residue 56 has significant influence on the activity of PLA2, and refute a recent report that substrate-level acylation of Lys-56 is an obligatory step in the catalysis by PLA2.
Localization of the second calcium ion binding site in porcine and equine phospholipase A2.
TLDR
Kinetic experiments on the various phospholipases finally demonstrated that enzyme species containing Glu71 bind a second Ca2+ ion to the low-affinity site, whereas enzymes lacking Glu 71 also lack this second site, confirming the suggestion that Glu72 is one of the ligands for Ca2- in the low/high affinity site.
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