Hayashi T, Iwai K, Ui N
Departments of Protein Chemistry and Physical Biochemistry, Institute of Endocrinology, Gunma University, Maebashi, Japan.
Biochim Biophys Acta. 1971 Nov 19;251(2):208-16. doi: 10.1016/0005-2795(71)90104-8.
In order to identify the number and types of peptide chains in thyroglobulin, noniodinated 19-S thyroglobulin obtained from goitrogen-treated hogs was exhaustively digested with trypsin (EC 3.4.4.4) after reduction and S-carboxymethylation. The digestion mixture was preliminarily separated into 30 fractions on Sephadex G-100 or G-15 and SE-Sephadex columns. The number of various tryptic peptides contained in each fraction was determined on peptide maps, where spots were detected with ninhydrin for total peptides and with each specific reagent for arginine, histidine or tyrosine-containing peptides. The number of total peptides observed in most of the fractions was estimated to be half the number of lysine plus arginine residues found in each fraction per mole of thyroglobulin, and the number of specific peptides was also close to half the number of each specific amino acid. These findings imply that thyroglobulin has 2-fold symmetry in the structure at the level of tryptic fragments and thus probably at the level of intact peptide chains.
为了确定甲状腺球蛋白中肽链的数量和类型,对从用致甲状腺肿物质处理过的猪中获得的非碘化19-S甲状腺球蛋白在还原和S-羧甲基化后用胰蛋白酶(EC 3.4.4.4)进行彻底消化。消化混合物在Sephadex G-100或G-15以及SE-Sephadex柱上初步分离成30个级分。在肽图上测定每个级分中所含各种胰蛋白酶肽的数量,在肽图上,用茚三酮检测总肽的斑点,用每种特定试剂检测含精氨酸、组氨酸或酪氨酸的肽的斑点。在大多数级分中观察到的总肽数量估计为每摩尔甲状腺球蛋白在每个级分中发现的赖氨酸加精氨酸残基数量的一半,并且特定肽的数量也接近每种特定氨基酸数量的一半。这些发现表明,甲状腺球蛋白在胰蛋白酶片段水平的结构上具有2倍对称性,因此可能在完整肽链水平上也具有2倍对称性。