Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis
Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis
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DOI：
10.1007/s003600050140
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发表时间：
1998-04-01
期刊：
影响因子：
--
通讯作者：
Parmelee, DC
中科院分区：
文献类型：
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作者：
Duman, JG;Li, N;Parmelee, DC
根据互补 DNA (cDNA) 和肽测序，确定了加拿大 Dendroides canadensis 甲虫幼虫的抗冻蛋白 (AFP) 推导的氨基酸序列。它们由具有 25 个残基信号肽的蛋白质和长度为 83 个（Dendroides 抗冻蛋白；DAFP-1）或 84 个（DAFP-2）氨基酸的成熟蛋白质组成，仅在两个位置上不同。肽测序产生的序列与推导的 DAFF-1 和 DAFP-2 的 cDNA 序列完全重叠，而另一个 AFP (DAFP-3) 的部分序列与 28 个残基中的 21 个匹配。这些抗冻蛋白中存在七个 12 或 13 聚体重复单元，其共有序列由以下组成：Cys-Thr-X-3-Ser-X-5-X-6-Cys-X-8-X-9-Ala-X-11-Thr-X-13，其中 X-3 和 X-11 倾向于带电残基，X-5 倾向于苏氨酸或丝氨酸，X-6 倾向于天冬酰胺或在 13 聚体中，X-9 朝向天冬酰胺或赖氨酸，X-13 朝向丙氨酸。这些蛋白质最有趣的特征是，在成熟抗冻蛋白的整个长度中，每六个残基就是一个半胱氨酸。这些序列与任何已知的鱼类 AFP 都不相似，但与黄粉虫的 AFP 相似。
The deduced amino acid sequences of antifreeze proteins (AFPs) from larvae of the beetle Dendroides canadensis were determined from both complementary DNAs (cDNAs) and from peptide sequencing. These consisted of proteins with a 25-residue signal peptide and mature proteins 83 (Dendroides antifreeze protein; DAFP-1) or 84 (DAFP-2) amino acids in length which differed at only two positions. Peptide sequencing yielded sequences which overlapped exactly with those of the deduced cDNA sequences of DAFF-1 and DAFP-2, while the partial sequence of another AFP (DAFP-3) matched 21 of 28 residues. Seven 12- or 13-mer repeating units are present in these antifreeze proteins with a consensus sequence consisting of: Cys-Thr-X-3-Ser-X-5-X-6-Cys-X-8-X-9-Ala-X-11-Thr-X-13, where X-3 and X-11 tend toward charged residues, X-5 tends toward threonine or serine, X-6 toward asparagine or aspartate, X-9 toward asparagine or lysine, and X-13 toward alanine in the 13-mers. The most interesting feature of these proteins is that throughout the length of the mature antifreeze proteins every sixth residue is a cysteine. These sequences are not similar to any of the known fish AFPs, but they are similar to AFPs from the beetle Tenebrio molitor.