Isolation and characterization of a novel perivitellin from the eggs of Pomacea scalaris (Mollusca, Ampullariidae).
Perivitellins are important components of the perivitelline fluid (PVF) that surrounds gastropod embryos. The glyco-lipo-carotenoprotein ovorubin (OR) from eggs of the snail Pomacea canaliculata has been the most studied to date. Here we report the characterization of scalarin (SC), a glyco-lipo-carotenoprotein from the PVF of P. scalaris. SC was purified by ultracentrifugation and exclusion chromatography. It is the major egg protein, representing 64% of the total soluble protein. The particle has a hydration density of 1.26 g/ml, an apparent molecular mass of 380 kDa and it is an elongated compact protein as estimated by small angle X-ray scattering (SAXS). It is composed of three subunits of ca. 35, 28, and 24 kDa noncovalently bonded. SC is highly glycosylated (carbohydrate content 20.1%, by wt.), with a low lipid content (0.7%), being esterified sterols, pigments and polar lipids the most abundant lipid classes. HPTLC and spectrophotometric analysis of the carotenoid fraction revealed the presence of free astaxanthin (ASX; 62.0%), and an unidentified carotenoid (38.0%). The carotenoid-apoprotein interaction was studied by spectrophotometry. Carotenoids do not seem to affect the structural characteristics of the oligomer. However, the carotenoid-protein association protected ASX against oxidation. The cross-reactivity between SC and perivitellins of P. canaliculata was tested using polyclonal antibodies (PAb) against SC, OR, and perivitellin PV2. The PAbs failed to cross-react with any egg proteins of either the same or other species. SC, among other functional similarities with OR, would be an antioxidant carrier, protecting at the same time carotenoids from oxidation in the perivitellin fluid of the egg.
Ituarte
S
S
Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CONICET-UNLP, La Plata, Argentina.
Dreon
M S
MS
Ceolín
M
M
Heras
H
H
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002338:Carotenoids / Q000032:analysis; D004527:Egg Proteins / Q000032:analysis* / Q000302:isolation & purification*; D005260:Female; D008055:Lipids / Q000032:analysis; D008970:Molecular Weight; D008974:Mollusca / Q000737:chemistry*; D018384:Oxidative Stress / Q000502:physiology; D021122:Protein Subunits / Q000032:analysis / Q000302:isolation & purification
D002241:Carbohydrates; D004527:Egg Proteins; D008055:Lipids; D021122:Protein Subunits; D002338:Carotenoids