Egorov T A, Odintsova T I, Musolyamov A K, Barbashov S F, Pustobaev V N, Andersen J, Roepstorff P, Popineau Y
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 117984, Russia.
Biochemistry (Mosc). 1998 Sep;63(9):1061-7.
We have determined the partial amino acid sequence (207 amino acids) of gamma-46 gliadin isolated from wheat cultivar Hardi. The molecular mass of the protein (Mr) estimated by electrospray mass spectrometry is 35191.3. The number of cysteine residues in gamma-46 gliadin was determined as a mass difference of the protein before and after reduction and alkylation with 4-vinylpyridine. It was shown that the protein has no free SH-groups, and all cysteine residues are involved in the formation of four disulfide bonds. The partial structure of gamma-46 gliadin was determined by N-terminal sequencing and sequencing of tryptic and chymotryptic peptides. The tryptic peptides were obtained by enzymatic hydrolysis of the protein, which was preliminarily reduced and immobilized at free SH-groups on thiopropyl-Sepharose 6B. The chymotryptic peptides were isolated by limited digestion of the native protein. The positions of cysteine residues, as well as surrounding amino acid sequences, are conserved in gamma-46 gliadin; this is typical of gliadins.
我们已经测定了从小麦品种Hardi中分离出的γ-46醇溶蛋白的部分氨基酸序列（207个氨基酸）。通过电喷雾质谱法估算的该蛋白质的分子量（Mr）为35191.3。γ-46醇溶蛋白中半胱氨酸残基的数量是通过用4-乙烯基吡啶进行还原和烷基化前后蛋白质的质量差异来确定的。结果表明该蛋白质没有游离的SH基团，所有半胱氨酸残基都参与形成四个二硫键。γ-46醇溶蛋白的部分结构通过N端测序以及胰蛋白酶和糜蛋白酶肽段的测序来确定。胰蛋白酶肽段是通过对蛋白质进行酶解获得的，该蛋白质预先进行了还原并固定在硫丙基-琼脂糖6B上的游离SH基团上。糜蛋白酶肽段是通过对天然蛋白质进行有限消化分离得到的。半胱氨酸残基的位置以及周围的氨基酸序列在γ-46醇溶蛋白中是保守的；这是醇溶蛋白的典型特征。
