Chandra T, Stackhouse R, Kidd V J, Robson K J, Woo S L
Biochemistry. 1983 Oct 25;22(22):5055-61. doi: 10.1021/bi00291a001.
alpha 1-Antichymotrypsin mRNA was isolated by specific polysome immunoprecipitation from turpentine-treated baboon liver. The highly enriched mRNA was used for synthesis and cloning of the corresponding cDNA. Baboon alpha 1-antichymotrypsin cDNA clones were identified by hybrid-selected translation, and the insert DNA fragment from one of the putative clones was used as a probe to screen a human liver cDNA library comprised of 40 000 independent transformants. One of the human cDNA clones was unambiguously identified to contain alpha 1-antichymotrypsin DNA sequences by comparison of its 5'-terminal nucleotide sequence with the N-terminal amino acid sequence of the protein. This cDNA clone, designated phACT235, contains 1524 base pairs of human DNA, which was sequenced in its entirety. The inserted DNA codes for a 25 amino acid signal peptide sequence and the entire mature alpha 1-antichymotrypsin of 408 amino acid residues. Comparison of the amino acid sequence of alpha 1-antichymotrypsin with that of the human alpha 1-antitrypsin has revealed a homology level similar to that between chymotrypsin and trypsin.
通过特异性多核糖体免疫沉淀法从松节油处理的狒狒肝脏中分离出α1-抗糜蛋白酶mRNA。高度富集的mRNA用于相应cDNA的合成和克隆。通过杂交选择翻译鉴定狒狒α1-抗糜蛋白酶cDNA克隆，并将其中一个推定克隆的插入DNA片段用作探针，筛选由40000个独立转化体组成的人肝脏cDNA文库。通过将其5'-末端核苷酸序列与该蛋白质的N-末端氨基酸序列进行比较，明确鉴定出一个人cDNA克隆含有α1-抗糜蛋白酶DNA序列。这个cDNA克隆命名为phACT235，包含1524个碱基对的人DNA，已对其进行了全序列测定。插入的DNA编码一个25个氨基酸的信号肽序列和由408个氨基酸残基组成的完整成熟α1-抗糜蛋白酶。α1-抗糜蛋白酶的氨基酸序列与人类α1-抗胰蛋白酶的氨基酸序列比较显示，其同源性水平与胰凝乳蛋白酶和胰蛋白酶之间的同源性水平相似。
