Kath T, Schmid R, Schäfer G
Institute for Biochemistry, Medical University of Lübeck, FRG.
Arch Biochem Biophys. 1993 Dec;307(2):405-10. doi: 10.1006/abbi.1993.1607.
An adenylate kinase gene from a member of the archaebacterial kingdom, the thermoacidophilic archaebacterium (archaeon) Sulfolobus acidocaldarius, has been cloned and sequenced for the first time. Two degenerate oligonucleotide probes, based on the N-terminal amino acid sequence information, led to the amplification of a gene-specific DNA fragment, used to screen subgenomic libraries. Comparing the DNA-derived amino acid sequence of total 194 residues with those of known procaryotic and eucaryotic adenylate kinases revealed only a low degree of similarity, except for a glycine-rich region close to the N-terminus, the so-called P-loop. Using inducible expression systems catalytically active S. acidocaldarius adenylate kinase was produced in large amounts. Although the total length of the protein and the results of alignment procedures suggest a closer relation to eucaryotic than to procaryotic sequences, the archaebacterial enzyme may represent a novel class of adenylate kinases. This is corroborated by the finding that an antiserum against this protein does not cross-react with Escherichia coli nor yeast or rabbit adenylate kinases for example.
首次克隆并测序了古细菌界嗜热嗜酸古细菌（古生菌）嗜酸热硫化叶菌的腺苷酸激酶基因。基于N端氨基酸序列信息设计的两个简并寡核苷酸探针，导致了一个基因特异性DNA片段的扩增，该片段用于筛选亚基因组文库。将总共194个残基的DNA衍生氨基酸序列与已知的原核和真核腺苷酸激酶的序列进行比较，发现除了靠近N端的富含甘氨酸的区域（即所谓的P环）外，相似性程度较低。使用诱导表达系统大量生产了具有催化活性的嗜酸热硫化叶菌腺苷酸激酶。尽管蛋白质的全长和比对程序结果表明该序列与真核序列的关系比与原核序列的关系更密切，但古细菌酶可能代表了一类新型的腺苷酸激酶。例如，针对该蛋白的抗血清与大肠杆菌、酵母或兔腺苷酸激酶不发生交叉反应，这一发现证实了这一点。
