HUMAN ALPHA-GALACTOSIDASE-A - NUCLEOTIDE-SEQUENCE OF A CDNA CLONE ENCODING THE MATURE ENZYME
HUMAN ALPHA-GALACTOSIDASE-A - NUCLEOTIDE-SEQUENCE OF A CDNA CLONE ENCODING THE MATURE ENZYME
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DOI：
10.1073/pnas.83.13.4859
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发表时间：
1986-07-01
影响因子：
11.1
通讯作者：
DESNICK, RJ
中科院分区：
文献类型：
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作者：
BISHOP, DF;CALHOUN, DH;DESNICK, RJ
已经确定了.lambda的完整核苷酸序列。gt11 cDNA克隆（.lambda. ag18）包含人α成熟溶酶体形式的全长编码区。-半乳糖苷酶A加一个;EC 3.2.1.22)。.lambda。AG18插入片段包含1226碱基对序列，开放阅读框编码成熟多肽的398个氨基酸（预测Mr = 45.356）和前肽序列的最后5个氨基酸。聚(A)信号AATACA和ATTAAA分别位于TAA终止密码子的28和11个核苷酸之前。由于poly(A)序列紧接在TAA终止密码子之后，因此没有3‘’未翻译区；第二个独立克隆的cDNA证实了这一发现。预测的氨基酸序列与86个不重叠残基（22%的成熟亚基）共线性，这些残基是通过对纯化的成熟酶的氨基端、色氨酸和溴化氰肽的微测序确定的。鉴定出4个潜在的n -糖基化位点，均发生在预测的。β。在二级结构的亲水性区域发生转动。HeLa poly(A)+ RNA的RNA转移杂交分析表明，其信号被单特异性的。α聚合体预先免疫吸收而降低。-Gal A抗体。核酸和蛋白质数据库的搜索没有发现显著的同源性，甚至与有限的序列可用于哺乳动物溶酶体酶。
The complete nucleotide sequence has been determined for a .lambda.gt11 cDNA clone (.lambda.AG18) containing the full-length coding region for the mature lysosomal form of human .alpha.-galactosidase A (.alpha.-Gal A; EC 3.2.1.22). The .lambda.AG18 insert contained a 1226-base-pair sequence with an open reading frame encoding 398 amino acids of the mature polypeptide (predicted Mr = 45.356) and the last 5 amino acids of the propeptide sequence. The poly(A) signals AATACA and ATTAAA occurred 28 and 11 nucleotide prior to the TAA stop codon, respectively. There was no 3'' untranslated region as the poly(A) sequence immediately followed the TAA termination codon; a second independently cloned cDNA confirmed this finding. The predicted amino acid sequence was colinear with 86 nonoverlapping residues (22% of the mature subunit) determined by microsequencing amino-termial, tryptic, and cyanogen bromide peptides of the purified mature enzyme. Four potential N-glycosylation sites were identified, all of which occurred at predicted .beta. turns in hydrophilic regions of seconary structure. RNA transfer hybridization analysis of HeLa poly(A)+ RNA demonstrated a single 1.45-kolobase band whose signal was decreased by prior immunoabsorption of polysomes with monospecific .alpha.-Gal A antibodies. Searches of nuclei acid and protein data bases did not reveal significant homology even with the limited sequences available for mammalian lysosomal enzymes.