Central Research and Development Department, E. I. du Pont de Nemours and Company, Wilmington, Delaware 19898.
Proc Natl Acad Sci U S A. 1982 Nov;79(22):6782-6. doi: 10.1073/pnas.79.22.6782.
The nitrogen fixation nif K gene of the blue-green alga Anabaena, which codes for the beta-subunit of dinitrogenase, has been subjected to sequence analysis. The nif K protein is predicted to be 512 amino acids long, to have a M(r) or 57,583, and to contain six cysteine residues. Three of these cysteines are within peptides homologous to FeS cluster-binding cysteinyl peptides from ferredoxins and from a high potential iron protein and, thus, may be ligands to which FeS clusters bind in dinitrogenase. The sequences surrounding the cysteine residues are 70% homologous to the corresponding cysteinyl tryptic peptides of the Azotobacter vinelandii dinitrogenase, although the positions of the cysteine residues are not always conserved between the two proteins. A 15-amino acid coding sequence precedes nif K on its transcript. Amino acid codon usage is highly asymmetric and parallels that found for the Anabaena dinitrogenase reductase gene (nif H). Putative promoter and ribosome binding site sequences were identified for nif K. These regulatory sequences are homologous to sequences preceding nif D; nif D codes for the alpha-subunit of dinitrogenase but is separated from nif K on the chromosome by 11,000 nucleotides. The nif K promoter also is virtually identical to a promoter-like sequence that immediately precedes the start of the transcript for the large subunit of ribulosebisphosphate carboxylase from maize chloroplasts. This homology appears to support the theory that chloroplasts evolved from blue-green algae.
蓝藻鱼腥藻固氮 nif K 基因的序列分析。该基因编码二氮酶的β亚基。nif K 蛋白预测由 512 个氨基酸组成,相对分子质量为 57583,含有 6 个半胱氨酸残基。其中 3 个半胱氨酸残基位于与铁氧还蛋白和高电位铁蛋白中 FeS 簇结合的半胱氨酸肽同源的肽段中,因此可能是二氮酶中 FeS 簇的配体。半胱氨酸残基周围的序列与固氮酶阿氏肠杆菌的相应半胱氨酰胰蛋白酶肽段有 70%的同源性,尽管两个蛋白质中半胱氨酸残基的位置并不总是保守的。nif K 转录物在其前面有 15 个氨基酸的编码序列。氨基酸密码子的使用高度不对称,与鱼腥藻二氮酶还原酶基因(nif H)的使用情况相似。nif K 鉴定了推定的启动子和核糖体结合位点序列。这些调节序列与 nif D 前面的序列同源;nif D 编码二氮酶的α亚基,但在染色体上与 nif K 隔开 11000 个核苷酸。nif K 启动子也与玉米叶绿体核酮糖二磷酸羧化酶大亚基转录物起始点前面的类似启动子序列几乎完全相同。这种同源性似乎支持了叶绿体从蓝藻进化而来的理论。