Molecular cloning of the sphingolipid activator protein-1 (SAP-1), the sulfatide sulfatase activator.
Molecular cloning of the sphingolipid activator protein-1 (SAP-1), the sulfatide sulfatase activator.
复制标题
鞘脂激活蛋白-1 (SAP-1)（硫苷脂硫酸酯酶激活剂）的分子克隆。
DOI：
10.1016/s0006-291x(86)80518-6
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发表时间：
1986
影响因子：
3.1
通讯作者：
O'Brien,JS
中科院分区：
文献类型：
--
作者：
Dewji,N;Wenger,D;Fujibayashi,S;Donoviel,M;Esch,F;Hill,F;O'Brien,JS
使用针对人SAP-1的多克隆抗体从λ gt 11人肝癌表达文库中分离编码SAP-1的cDNA。分离到三个阳性克隆，插入片段约为0.3 Kb（S1.1）、2 Kb（S1.2）和2.2 Kb（S-1.3）。后2个含有一个内部的EcoRI位点。所有三个克隆彼此交叉杂交，表明序列同源性。测定了S-1.1的核苷酸序列。通过对纯SAP-1的氨基末端测序获得的19个氨基酸与S-1.1的5′和57 bp之间建立了共线性。S-1.1的开放阅读框编码67个氨基酸。在离氨基末端21个残基处发现一个糖基化位点，未发现终止密码子。S-1.1编码成熟多肽链，计算分子量为8955道尔顿，相当于成熟SAP-1的约99%。
A cDNA coding for SAP-1 was isolated from a λgt11 human hepatoma expression library using polyclonal antibodies raised against human SAP-1. Three positive clones were isolated with inserts of approximately 0.3 Kb (S1.1), 2 Kb (S1.2) and 2.2 Kb (S-1.3). The latter 2 contained an internal EcoRI site. All three clones cross-hybridized with one another, indicating sequence homology. The nucleotide sequence of S-1.1 was determined. Colinearity was established between 19 amino acids obtained by sequencing the amino terminus of pure SAP-1 and 57 bp from the 5′ and of S-1.1. The open reading frame of S-1.1 coded for 67 amino acids. One glycosylation site was found 21 residues from the amino terminus, and no stop codons were found. S-1.1 codes for a mature polypeptide chain with a calculated molecular weight of 8955 daltons, corresponding to approximately 99% of mature SAP-1.