Metabolism of ovorubin, the major egg lipoprotein from the apple snail

The site of synthesis of molluscs lipoproteins is little known and was investigated for the egg lipoprotein perivitellin 1 (PV 1) or ovorubin in the freshwater snail Pomacea canaliculata. Tissues (albumen gland, gonad-digestive gland complex and muscle) of vitellogenic females were incubated in vitr...

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Bibliographic Details
Published in:Molecular and cellular biochemistry 2003-01, Vol.243 (1-2), p.9-14
Main Authors: Dreon, Marcos S, Heras, Horacio, Pollero, Ricardo J
Format: Article
Language:English
Subjects:
Animals
Bacteria
Blotting, Western
Chromatography, High Pressure Liquid
Digestive glands
Egg Proteins - chemistry
Egg Proteins - metabolism
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Female
Freshwater
Immunoblotting
Leucine - chemistry
Lipoproteins - chemistry
Liquid chromatography
Mollusca
Mollusks
Pomacea canaliculata
Protein synthesis
Proteins
Snails
Temperature
Time Factors
Tissue Distribution
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Summary:The site of synthesis of molluscs lipoproteins is little known and was investigated for the egg lipoprotein perivitellin 1 (PV 1) or ovorubin in the freshwater snail Pomacea canaliculata. Tissues (albumen gland, gonad-digestive gland complex and muscle) of vitellogenic females were incubated in vitro at 25 degrees C for 12 h with 14C Leucine. After that, soluble proteins from tissue homogenates and medium samples were analysed for de novo protein synthesis by electrophoresis and HPLC, and radiolabelled proteins quantified by liquid scintillation. Gonad-digestive gland complex did not synthesise ovorubin, in spite its high protein synthesis levels. Three albumen gland radiolabelled proteins (35, 32 and 28 kDa) comigrated with the subunits of ovorubin and represented 1.3% of the total labelled protein of that tissue. Western blot analysis with polyclonal antibodies confirmed that these were ovorubin subunits. In vivo experiments where vitellogenic females were injected with 3H Leucine, revealed that ovorubin was not present in hemolymph. ELISA analysis confirmed ovorubin presence only in albumen gland and developing eggs with levels of 800 and 582 mg/g protein, which represent 30.3 and 28.4 mg ovorubin/g of tissue, respectively. Therefore, albumen gland is the single site of ovorubin synthesis as no extragland synthesis, circulation or accumulation could be detected in the apple snail.
ISSN:0300-8177
1573-4919
DOI:10.1023/A:1021616610241