The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae. Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes.
The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae. Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes.
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DOI：
10.1016/s0021-9258(19)81564-8
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发表时间：
1988-07
期刊：
影响因子：
--
通讯作者：
E. Schwarz;D. Oesterhelt;H. Reinke;K. Beyreuther;P. Dimroth
中科院分区：
文献类型：
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作者：
E. Schwarz;D. Oesterhelt;H. Reinke;K. Beyreuther;P. Dimroth
克隆并测序了编码肺炎克雷伯菌Na+泵草乙酸脱羧酶α亚基的基因。通过约30%的多肽链的蛋白质测序证实了推导的蛋白质一级结构。该基因GC含量为67%，编码596个氨基酸。成熟蛋白中的 N 端甲硫氨酸被去除，计算得出的分子量为 63,600 道尔顿。该蛋白质由两个不同的结构域组成，这两个结构域通过一段易于蛋白水解切割的氨基酸残基连接。用胰蛋白酶对天然酶进行有限的蛋白水解，产生约 51 kDa 和 10.2 kDa 的片段，后者从缬氨酸 491 开始并包含生物素辅基。肽测序表明生物素辅基与 561 位赖氨酸（距 C 末端 35 个残基）结合。脱羧酶在 10.2-kDa 生物素结构域的 N 端侧包含延伸的富含丙氨酸和脯氨酸的区域（位置 502-532）。该序列总共包括 16 个丙氨酸和 9 个脯氨酸残基。
The gene encoding the alpha-subunit of the Na+ pump oxalacetate decarboxylase of Klebsiella pneumoniae was cloned and sequenced. The deduced primary structure of the protein was confirmed by protein sequencing of about 30% of the polypeptide chain. The gene has a GC content of 67% and codes for 596 amino acids. The N-terminal methionine is removed in the mature protein which has a calculated molecular mass of 63,600 daltons. The protein consists of two different domains that are connected by a stretch of amino acid residues susceptible to proteolytic cleavage. Limited proteolysis of the native enzyme with trypsin produced fragments of about 51 kDa and 10.2 kDa, the latter of which started with valine 491 and contained the biotin prosthetic group. Peptide sequencing indicated binding of the biotin prosthetic group to lysine 561, 35 residues from the C terminus. The decarboxylase contains an extended alanine- and proline-rich region (positions 502-532) on the N-terminal side of the 10.2-kDa biotin domain. This sequence includes a total of 16 alanine and 9 proline residues.