Nonaka M, Takahashi M, Natsuume-Sakai S, Nonaka M, Tanaka S, Shimizu A, Honjo T
Proc Natl Acad Sci U S A. 1984 Nov;81(21):6822-6. doi: 10.1073/pnas.81.21.6822.
cDNA clones specific for the fourth component of mouse complement (C4) and its hormonally regulated isotype, sex-linked protein (Slp), were isolated using as a probe a 20-mer synthetic oligonucleotide corresponding to a known sequence of human C4 cDNA. Two types of clones, one specific for C4 (pFC4/10, with a 3.7 kilobase insert) and one specific for Slp (pFSlp/1, with a 4.7 kilobase insert), were isolated from liver cDNA libraries constructed from the Slp-producing FM mouse strain. The cDNA inserts of these clones shared 70% of the restriction sites determined. Only one type of clone was isolated from the Slp-negative DBA/1 strain; this type showed restriction maps indistinguishable from that of pFC4/10. pFC4/10 and pFSlp/1 displayed extensive homology: 94% nucleotide homology and 89% derived amino acid homology in the C4a region and 92% nucleotide homology and 89% derived amino acid homology in the thiol-ester region. An Arg-Gln-Lys-Arg sequence in the beta-alpha junction and a Cys-Ala-Glu-Gln sequence in the thiol-ester site were identified for both proteins. A remarkable divergency between C4 and Slp sequences was recognized in the region immediately following the C4a sequence.
利用一段与人类C4 cDNA已知序列相对应的20聚体合成寡核苷酸作为探针,分离出了小鼠补体第四成分(C4)及其激素调节同种型——性连锁蛋白(Slp)的特异性cDNA克隆。从由产生Slp的FM小鼠品系构建的肝脏cDNA文库中,分离出了两种类型的克隆,一种对C4特异(pFC4/10,插入片段为3.7千碱基),另一种对Slp特异(pFSlp/1,插入片段为4.7千碱基)。这些克隆的cDNA插入片段共有70%已确定的限制性酶切位点。从Slp阴性的DBA/1品系中只分离出了一种类型的克隆;这种类型的限制性图谱与pFC4/10的无法区分。pFC4/10和pFSlp/1显示出广泛的同源性:在C4a区域核苷酸同源性为94%,推导氨基酸同源性为89%;在硫酯区域核苷酸同源性为92%,推导氨基酸同源性为89%。两种蛋白质在β-α连接区均鉴定出一个精氨酸-谷氨酰胺-赖氨酸-精氨酸序列,在硫酯位点均鉴定出一个半胱氨酸-丙氨酸-谷氨酸-谷氨酰胺序列。在紧接C4a序列之后的区域,C4和Slp序列之间存在显著差异。