Structure of the molybdopterin-bound Cnx1G domain links molybdenum and copper metabolism.
The molybdenum cofactor is part of the active site of all molybdenum-dependent enzymes, except nitrogenase. The molybdenum cofactor consists of molybdopterin, a phosphorylated pyranopterin, with an ene-dithiolate coordinating molybdenum. The same pyranopterin-based cofactor is involved in metal coordination of the homologous tungsten-containing enzymes found in archea. The molybdenum cofactor is synthesized by a highly conserved biosynthetic pathway. In plants, the multidomain protein Cnx1 catalyses the insertion of molybdenum into molybdopterin. The Cnx1 G domain (Cnx1G), whose crystal structure has been determined in its apo form, binds molybdopterin with high affinity and participates in the catalysis of molybdenum insertion. Here we present two high-resolution crystal structures of Cnx1G in complex with molybdopterin and with adenylated molybdopterin (molybdopterin-AMP), a mechanistically important intermediate. Molybdopterin-AMP is the reaction product of Cnx1G and is subsequently processed in a magnesium-dependent reaction by the amino-terminal E domain of Cnx1 to yield active molybdenum cofactor. The unexpected identification of copper bound to the molybdopterin dithiolate sulphurs in both structures, coupled with the observed copper inhibition of Cnx1G activity, provides a molecular link between molybdenum and copper metabolism.
Department of Plant Biology, Technical University, Spielmannstrasse 7, D-38106 Braunschweig, Germany.
Jochen
J
Kuper
Angel
A
Llamas
Hans-Jürgen
HJ
Hecht
Ralf R
RR
Mendel
Günter
G
Schwarz
D000249:Adenosine Monophosphate / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D037281:Calnexin / Q000737:chemistry / Q000378:metabolism; D003067:Coenzymes / Q000378:metabolism*; D003300:Copper / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008274:Magnesium / Q000378:metabolism; D008667:Metalloproteins / Q000378:metabolism*; D008958:Models, Molecular; D008982:Molybdenum / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism*; D010944:Plants / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011621:Pteridines / Q000378:metabolism*; D013455:Sulfur / Q000378:metabolism
D000249:Adenosine Monophosphate; D029681:Arabidopsis Proteins; D037281:Calnexin; D003067:Coenzymes; D003300:Copper; D008274:Magnesium; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D010940:Plant Proteins; D011621:Pteridines; D013455:Sulfur; C080351:CNX1 protein, Arabidopsis; C022261:molybdopterin