Salerno Ana Paula, Dansa-Petretski Marílvia, Silva-Neto Mário A C, Coelho Heloísa S L, Masuda Hatisaburo
Departamento de Bioquímica Médica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21940-590, Brazil.
Insect Biochem Mol Biol. 2002 Jul;32(7):709-17. doi: 10.1016/s0965-1748(01)00152-7.
Rhodnius prolixus oocyte extracts were chromatographed on an ion exchange column in order to purify vitellin (VT). Three VT heterogeneous populations were identified and named VT(1), VT(2), and VT(3) according to their order of elution from the column. The phosphate content of each population was determined, after lipid extraction, and a heterogeneous distribution was found: VT(1) being the less phosphorylated (50 mol P/mol protein) and VT(3) the heavily phosphorylated population (281 mol P/mol protein). Analysis of radioactivity associated with each VT population purified from animals fed with (32)Pi showed the same phosphorylation profile. Due to the fact that vitellogenin is the known precursor of VT, we have also chromatographed 32P-VG in the same way as we purified VT. Only one VG's population was detected and resembled to VT(3) with respect to its elution profile. All VT populations contain the same neutral lipids, but they were heterogeneous with respect to phospholipid composition. VT(1) presents phosphatidylcholine and phosphatidylethanolamine whereas VT(2) and VT(3) also showed cardiolipin and probably phosphatidylserine. Sugar composition of VT(2) and VT(3) includes mannose as the main associated carbohydrate but VT(1) also contains glucose resembling VG. Although VG and VT are similar with respect to the elution profile, their sugar composition is different. These results suggest a post-endocytosis processing on VG molecule. The possible biological function of VT heterogeneous populations is discussed.
为了纯化卵黄磷蛋白(VT),将红带锥蝽的卵母细胞提取物在离子交换柱上进行色谱分析。根据它们从柱上洗脱的顺序,鉴定出了三个VT异质群体,并分别命名为VT(1)、VT(2)和VT(3)。在脂质提取后,测定了每个群体的磷酸盐含量,发现其分布不均:VT(1)的磷酸化程度最低(50摩尔磷/摩尔蛋白质),而VT(3)是磷酸化程度高的群体(281摩尔磷/摩尔蛋白质)。对从用(32)Pi喂养的动物中纯化得到的每个VT群体相关放射性的分析显示出相同的磷酸化谱。由于卵黄蛋白原是已知的VT前体,我们也以纯化VT的相同方式对32P-VG进行了色谱分析。仅检测到一个VG群体,就其洗脱谱而言与VT(3)相似。所有VT群体都含有相同的中性脂质,但它们在磷脂组成方面是异质的。VT(1)含有磷脂酰胆碱和磷脂酰乙醇胺,而VT(2)和VT(3)还显示出心磷脂,可能还含有磷脂酰丝氨酸。VT(2)和VT(3)的糖组成包括甘露糖作为主要相关碳水化合物,但VT(1)还含有类似于VG的葡萄糖。尽管VG和VT在洗脱谱方面相似,但它们的糖组成不同。这些结果表明在VG分子上存在内吞后加工过程。本文讨论了VT异质群体可能的生物学功能。