Subunit composition of the zinc proteins alpha- and beta-lipovitellin from chicken.
Chicken alpha- and beta-lipovitellin are derived from parent vitellogenin proteins and contain four subunits (125, 80, 40, and 30 kDa) and two subunits (125 and 30 kDa), respectively. Metal analyses demonstrate both are zinc proteins containing 2.1 +/- 0.2 mol of zinc/275 kDa per alpha-lipovitellin and 1.4 +/- 0.2 mol of zinc/155 kDa per beta-lipovitellin, respectively. The subunits of beta-lipovitellin, Lv 1 (MW 125 kDa) and Lv 2 (MW 30 kDa), are separated by gel exclusion chromatography in the presence of zwittergent 3-16. Zinc elutes with Lv 1, suggesting that this subunit binds zinc in the absence of Lv 2. The subunits of alpha- and beta-lipovitellin were separated by SDS-PAGE, digested with trypsin, and mapped by reverse-phase HPLC. The peptide maps of the 125-kDa subunits from alpha- and beta-lipovitellin are essentially identical. Similar results are obtained for the 30-kDa subunits of both lipovitellins. The sequences of five and four peptides of the 125-kDa subunit of alpha- and beta-Lv, respectively, and two peptides of the 30-kDa subunit of alpha- and beta-lipovitellin were determined and match those predicted from the gene for vitellogenin II, Vtg II. Comparison of the amino acid composition of the 125- and 30-kDa subunits of alpha- and beta-lipovitellin support the conclusion that they originate from the same gene. The sequences of peptides from the 80- and 40-kDa subunits of alpha-lipovitellin have not been found in the NCBI nonredundant data bank. The 27-amino acid N-terminal sequence of the 40-kDa protein is 56% similar to the last third of the Lv 1-coding region of the Vtg II gene, suggesting it may come from an analogous region of the Vtg I gene. We propose a scheme for the precursor-product relationship of Vtg I.
Groche
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Center for Biochemical and Biophysical Sciences and Medicine, Brigham and Women's Hospital, Boston, Massachusetts, USA.
Rashkovetsky
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LG
Falchuk
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KH
Auld
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D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002645:Chickens; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004527:Egg Proteins; D015387:Egg Proteins, Dietary / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D010449:Peptide Mapping; D017386:Sequence Homology, Amino Acid; D014982:Xenopus laevis; D015032:Zinc / Q000032:analysis*
D000596:Amino Acids; D004527:Egg Proteins; D015387:Egg Proteins, Dietary; C001549:lipovitellin; D015032:Zinc