Lipocalin-type prostaglandin D synthase (beta-trace) is a newly recognized type of retinoid transporter.
Lipocalin-type prostaglandin D synthase is responsible for the biosynthesis of prostaglandin D2 in the central nervous system and the genital organs and is secreted into the cerebrospinal fluid and the seminal plasma as beta-trace. Here we analyzed retinoids binding of the enzyme by monitoring the fluorescence quenching of an intrinsic tryptophan residue, and appearance of circular dichroism around 330 nm, and a red shift of the UV absorption spectra of retinoids. We found that the enzyme binds all-trans- or 9-cis-retinoic acid and all-trans- or 13-cis-retinal, but not all-trans-retinol, with affinities (Kd of 70-80 nM) sufficient for function as a retinoid transporter. All-trans-retinoic acid inhibited the enzyme activity in a noncompetitive manner, suggesting that it binds to the same hydrophobic pocket as prostaglandin H2, the substrate for prostaglandin D synthase, but at a different site in this pocket. It is likely that this enzyme is a bifunctional protein that acts as both retinoid transporter and prostaglandin D2-producing enzyme.
Tanaka
T
T
Protein Engineering Research Institute, 6-2-3 Furuedai, Suita, Osaka 565, Japan.ttanaka@beri.co.jp
Urade
Y
Y
Kimura
H
H
Eguchi
N
N
Nishikawa
A
A
Hayaishi
O
O
D000409:Alanine / Q000378:metabolism; D000077556:Alitretinoin; D000818:Animals; D001693:Biological Transport, Active; D002942:Circular Dichroism; D003553:Cystine / Q000378:metabolism; D004224:Diterpenes; D019746:Intramolecular Oxidoreductases; D007535:Isomerases / Q000737:chemistry / Q000378:metabolism*; D007536:Isomerism; D007700:Kinetics; D054834:Lipocalins; D008958:Models, Molecular; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012172:Retinaldehyde / Q000378:metabolism; D012176:Retinoids / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000378:metabolism; D054839:Retinol-Binding Proteins, Plasma; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014212:Tretinoin / Q000378:metabolism
D004224:Diterpenes; D054834:Lipocalins; D011994:Recombinant Proteins; D012176:Retinoids; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D000077556:Alitretinoin; C031338:13-cis-retinal; D003553:Cystine; D014212:Tretinoin; D007535:Isomerases; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase; D000409:Alanine; D012172:Retinaldehyde