Molecular characterization of vitellin from the ovaries of the white shrimp Penaeus (Litopenaeus) vannamei.
Vitellin (Vt) was purified from ovary extracts of mature females of the white shrimp Penaeus vannamei using Sepharose CL-4B and Q-Sepharose columns. Native Vt had an apparent molecular weight of 388 kDa as detected in Native-PAGE, bound the lipophilic dye Oil Red O and had a total lipid content of approximately 43.8%. Under reducing and denaturing conditions (SDS-PAGE), Vt is composed of three major subunits of 87, 78 and 46 kDa, although minor bands of 65, 61 and 31 kDa are also detected. The 87- and 78-kDa polypeptides were strongly recognized by Penaeus semisulcatus anti-Vt polyclonal and Penaeus monodon anti-Vt monoclonal antibodies. Furthermore, the N-terminal amino acid sequence of the 78-kDa polypeptide is very similar to Penaeus japonicus vitellogenin (Vg) and P. semisulcatus Vt, with an identity of 76%. Circular dichroism indicates that the beta-helix content of Vt is 25% while beta-sheets correspond to 37 and 14% of unordered secondary structure. These values are similar to insect microvitellogenin. Vt has an emission fluorescence maximum at 329 nm, comparable to the shrimp high-density lipoprotein/beta-glucan binding protein (HDL/BGBP).
García-Orozco
Karina D
KD
Marine Biotechnology, Centro de Investigación en Alimentación y Desarrollo, PO Box 1735 Hermosillo Son, 83000 Mexico.
Vargas-Albores
Francisco
F
Sotelo-Mundo
Rogerio R
RR
Yepiz-Plascencia
Gloria
G
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004527:Egg Proteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D005260:Female; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010053:Ovary / Q000737:chemistry*; D033561:Penaeidae / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017385:Sequence Homology; D013050:Spectrometry, Fluorescence; D014819:Vitellogenins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification
D004527:Egg Proteins; D014819:Vitellogenins