Expression, purification, and characterization of NosL, a novel Cu(I) protein of the nitrous oxide reductase (nos) gene cluster.
NosL, one of the accessory proteins of the nos (nitrous oxide reductase) gene cluster, has been heterologously expressed, purified, and characterized. NosL is a monomeric protein of 18,540 MW that specifically and stoichiometrically binds Cu(I). The copper ion in NosL is ligated by a Cys residue, and one Met and one His are thought to serve as the other ligands. While it is possible to oxidize Cu(I)-NosL with ferricyanide, the Cu(II) ion thus formed appears to dissociate from the protein. The function of Cu(I)NosL is not yet known, but the data indicate that NosL does not act as an electron transfer partner to nitrous oxide reductase. NosL is encoded on the same transcript as three other gene products (NosD, NosF, and NosY). These have been shown to be required for assembly of the active site in nitrous oxide reductase, which is thought to be a copper cluster. Accordingly, it is possible that NosL is a copper chaperone involved in metallocenter assembly.
McGuirl
M A
MA
Department of Chemistry, and Biochemistry, Montana State University, Bozeman 59717, USA.
Bollinger
J A
JA
Cosper
N
N
Scott
R A
RA
Dooley
D M
DM
D000421:Alcaligenes / Q000737:chemistry*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D003300:Copper / Q000737:chemistry*; D004926:Escherichia coli; D005786:Gene Expression Regulation; D008074:Lipoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008667:Metalloproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones; D015394:Molecular Structure; D010088:Oxidoreductases; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013052:Spectrometry, X-Ray Emission
D001425:Bacterial Outer Membrane Proteins; D008074:Lipoproteins; D008667:Metalloproteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D003300:Copper; D010088:Oxidoreductases; C022763:nitrous oxide reductase