title: In vitro stimulatory effect of anti-apoptotic seminal vesicle protein 4 on purified peroxidase enzymes.
abstract: The enzymatic activities of purified horseradish peroxidase, selenium-dependent glutathione peroxidase, thyroid peroxidase and myeloperoxidase, but not that of lactoperoxidase, were markedly enhanced when added into a reaction mixture containing 5 mum native seminal vesicle protein 4, a major protein secreted from rat seminal vesicle epithelium. A further increase of horseradish peroxidase activity was obtained using Ser58-phosphorylated or acetylated seminal vesicle protein 4. The activating effect of native seminal vesicle protein 4 was highest (about 60-fold) on horseradish peroxidase when 4-chloro-1-naphtol was used as the electron donor substrate. The main kinetics parameters of the stimulatory effect on horseradish peroxidase were evaluated and the enzyme-electron donor substrate interaction was investigated by HPLC and electrospray-MS. A native seminal vesicle protein 4/4-chloro-1-naphtol noncovalent adduct was detected when the protein and 4-chloro-1-naphtol were present in the appropriate molar ratio in the horseradish peroxidase-catalyzed reaction. By contrast, no adducts were formed between native seminal vesicle protein 4 and horseradish peroxidase. This native seminal vesicle protein 4/4-chloro-1-naphtol interaction might underlie the native seminal vesicle protein 4-induced horseradish peroxidase stimulation. Furthermore, native seminal vesicle protein 4 was shown by spectrophotometric and electrospray-MS analysis to interact with NADPH, an electron donor substrate of the selenium-dependent glutathione peroxidase/glutathione reductase redox system, with formation of an adduct between them. Although further investigation is required to elucidate the mechanism of adduct formation, this interaction, probably by promoting the release of the NADPH electrons required for glutathione disulphide reduction, could explain the stimulatory effect of seminal vesicle protein 4 on mammalian peroxidases possibly involved in its physiological function on the selenium-dependent glutathione peroxidase/glutathione reductase system. The biological significance of these properties of native seminal vesicle protein 4 might be related to its ability to downregulate reactive oxygen species and oxidative stress-induced apoptosis.
authors.affiliation: Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", CNR, Naples, Italy. metafora@igb.cnr.it
authors.forename: Vittoria
authors.initials: V
authors.lastname: Metafora
authors.forename: Paola
authors.initials: P
authors.lastname: Stiuso
authors.forename: Pasquale
authors.initials: P
authors.lastname: Ferranti
authors.forename: Angela
authors.initials: A
authors.lastname: Giannattasio
authors.forename: Alessandra
authors.initials: A
authors.lastname: Dicitore
authors.forename: Gianpietro
authors.initials: G
authors.lastname: Ravagnan
authors.forename: Salvatore
authors.initials: S
authors.lastname: De Maria
authors.forename: Gabriele
authors.initials: G
authors.lastname: Pontoni
authors.forename: Maria
authors.initials: M
authors.lastname: Cartenì
authors.forename: Salvatore
authors.initials: S
authors.lastname: Metafora
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D066298:In Vitro Techniques; D008297:Male; D009249:NADP / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D051381:Rats; D029607:Seminal Vesicle Secretory Proteins / Q000378:metabolism / Q000502:physiology*; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet
chemical_list: D029607:Seminal Vesicle Secretory Proteins; C101178:Svs4 protein, rat; D009249:NADP; D010544:Peroxidases