Purification and structural determination of a phosphorylated peptide with anti-calcification and chitin-binding activities in the exoskeleton of the crayfish, Procambarus clarkii
Purification and structural determination of a phosphorylated peptide with anti-calcification and chitin-binding activities in the exoskeleton of the crayfish, Procambarus clarkii
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DOI：
10.1271/bbb.65.1840
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发表时间：
2001-08-01
影响因子：
1.6
通讯作者：
Nagasawa, H
中科院分区：
文献类型：
--
作者：
Inoue, H;Ozaki, N;Nagasawa, H
钙化硬组织中的有机基质被认为是控制钙化的因素。从克氏原螯虾（Procambarus clarkii）的外骨骼中提取并通过阴离子交换和反相高效液相色谱法纯化了一种基质肽CAP-1。对CAP-1的完整肽段及其氨基酸序列进行了质谱和序列分析。酶消化的肽。CAP-1由78个氨基酸残基组成，包括磷酸丝氨酸残基，并且富含酸性氨基酸残基。CAP-1具有Rebers-Riddiford共有序列，该序列在许多节肢动物的角质层蛋白中是保守的。CAP-1在体外抗钙化试验中剂量依赖性地抑制碳酸钙沉淀，在3 × 10 - 7时完全抑制。CAP-1还显示出几丁质结合能力，表明该分子是双功能的，并且在外骨骼的形成中起重要作用。
Organic matrices in calcified hard tissues have been considered to control calcification. A matrix peptide, designated CAP-1, was extracted and purified by anion-exchange and reverse-phase high performance liquid chromatographies from the exoskeleton of the crayfish, Procambarus clarkii. The amino acid sequence of CAP-1 was determined by mass spectral and sequence analyses of the intact peptide and its. enzymatically digested peptides. CAP-1 consisted of 78 amino acid residues, including a phosphoserine residue, and was rich in acidic amino acid residues. CAP-1 had a Rebers-Riddiford consensus sequence, which is conserved in cuticle proteins from many arthropods. CAP-1 inhibited precipitation of calcium carbonate in an in vitro anticalcification assay dose-dependently, and completely inhibited it at 3 x 10(-7) :ii. CAP-1 also showed chitin-binding ability, indicating that this molecule was bifunctional and played an important role in formation of the exoskeleton.