Sulfhydryl oxidase from egg white. A facile catalyst for disulfide bond formation in proteins and peptides.
Both metalloprotein and flavin-linked sulfhydryl oxidases catalyze the oxidation of thiols to disulfides with the reduction of oxygen to hydrogen peroxide. Despite earlier suggestions for a role in protein disulfide bond formation, these enzymes have received comparatively little general attention. Chicken egg white sulfhydryl oxidase utilizes an internal redox-active cystine bridge and a FAD moiety in the oxidation of a range of small molecular weight thiols such as glutathione, cysteine, and dithiothreitol. The oxidase is shown here to exhibit a high catalytic activity toward a range of reduced peptides and proteins including insulin A and B chains, lysozyme, ovalbumin, riboflavin-binding protein, and RNase. Catalytic efficiencies are up to 100-fold higher than for reduced glutathione, with typical K(m) values of about 110-330 microM/protein thiol, compared with 20 mM for glutathione. RNase activity is not significantly recovered when the cysteine residues are rapidly oxidized by sulfhydryl oxidase, but activity is efficiently restored when protein disulfide isomerase is also present. Sulfhydryl oxidase can also oxidize reduced protein disulfide isomerase directly. These data show that sulfhydryl oxidase and protein disulfide isomerase can cooperate in vitro in the generation and rearrangement of native disulfide pairings. A possible role for the oxidase in the protein secretory pathway in vivo is discussed.
Hoober
K L
KL
Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware 19716, USA.
Sheasley
S L
SL
Gilbert
H F
HF
Thorpe
C
C
D000818:Animals; D002645:Chickens; D004220:Disulfides / Q000378:metabolism*; D004529:Egg White; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D005420:Flavoproteins / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D050862:Oxidoreductases Acting on Sulfur Group Donors
D004220:Disulfides; D005182:Flavin-Adenine Dinucleotide; D005420:Flavoproteins; D009113:Muramidase; D010088:Oxidoreductases; D010455:Peptides; D019704:Protein Disulfide-Isomerases; D011506:Proteins; D013438:Sulfhydryl Compounds; D050862:Oxidoreductases Acting on Sulfur Group Donors