The 1.23 Angstrom structure of Pichia pastoris lysyl oxidase reveals a lysine-lysine cross-link.
The structure of Pichia pastoris lysyl oxidase (PPLO) in a new crystal form has been refined at 1.23 Angstrom resolution. PPLO, a copper amine oxidase (CuAO) with a 2,4,5-trihydroxyphenylalanine quinone (TPQ) cofactor, differs from most other members of the CuAO enzyme family in having the ability to oxidize the side chain of lysine residues in a polypeptide. In the asymmetric unit of the crystals, the structure analysis has located residues 43-779 of the polypeptide chain, seven carbohydrate residues, the active-site Cu atom, an imidazole molecule bound at the active site, two buried Ca(2+) ions, five surface Mg(2+) ions, five surface Cl(-) ions and 1045 water molecules. The crystallographic residuals are R = 0.112 and R(free) = 0.146. The TPQ cofactor and several other active-site residues are poorly ordered, in contrast to the surrounding well ordered structure. A covalent cross-link is observed between two lysine residues, Lys778 and Lys66. The cross-link is likely to have been formed by the oxidation of Lys778 followed by a spontaneous reaction with Lys66. The link is modelled as dehydrolysinonorleucine.
Duff
Anthony P
AP
School of Molecular and Microbial Biosciences, University of Sydney, NSW 2006, Australia.
Cohen
Aina E
AE
Ellis
Paul J
PJ
Hilmer
Kim
K
Langley
David B
DB
Dooley
David M
DM
Freeman
Hans C
HC
Guss
J Mitchell
JM
D001665:Binding Sites; D003300:Copper / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018360:Crystallography, X-Ray / Q000379:methods*; D004151:Dipeptides / Q000737:chemistry; D006031:Glycosylation; D007477:Ions; D008239:Lysine / Q000737:chemistry*; D008956:Models, Chemical; D008958:Models, Molecular; D008968:Molecular Conformation; D010843:Pichia / Q000201:enzymology*; D011487:Protein Conformation; D008249:Protein-Lysine 6-Oxidase / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004151:Dipeptides; D007477:Ions; C007238:dehydrolysylnorleucine; D003300:Copper; D008249:Protein-Lysine 6-Oxidase; D008239:Lysine