Sandbox Reserved 1096: Difference between revisions
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To obtain the structure of '''''PAD2 holoenzyme''''', a double mutant F221/F222A was engineered to prevent undesired interactions with neighboring hydrophobic pockets that could prevent binding of Calcium at Ca2 site. Structure of PAD2 F221/222A Ca2+ mutant was solved and studied showing an important electron density at all 6 calcium binding sites at 10mM Ca2+. All sites are now binding with calcium. Moreover, The active site cysteine, C647 points toward the catalytic center making the holoenzyme competent for catalysis. Calcium binding to Ca2 site was also found to cause R347 to move out of the active site while W348 moves in to form one wall of the substrate-binding pocket. | To obtain the structure of '''''PAD2 holoenzyme''''', a double mutant F221/F222A was engineered to prevent undesired interactions with neighboring hydrophobic pockets that could prevent binding of Calcium at Ca2 site. Structure of PAD2 F221/222A Ca2+ mutant was solved and studied showing an important electron density at all 6 calcium binding sites at 10mM Ca2+. All sites are now binding with calcium. Moreover, The active site cysteine, C647 points toward the catalytic center making the holoenzyme competent for catalysis. Calcium binding to Ca2 site was also found to cause R347 to move out of the active site while W348 moves in to form one wall of the substrate-binding pocket. | ||
[[Image:PAD2.jpg|300px|right|thumb| Main transformations of apo and holo-enzymes that explain PAD2's activity]] | |||
==='''Catalysis of deimination'''=== | ==='''Catalysis of deimination'''=== | ||
