Centre Scientifique de Monaco, Avenue Saint-Martin, MC-98000, Monaco, France.
J Biol Chem. 2012 Jun 1;287(23):19367-76. doi: 10.1074/jbc.M112.352005. Epub 2012 Apr 13.
We report here for the first time the isolation and characterization of a protein from the organic matrix (OM) of the sclerites of the alcyonarian, Corallium rubrum. This protein named scleritin is one of the predominant proteins extracted from the EDTA-soluble fraction of the OM. The entire open reading frame (ORF) was obtained by comparing amino acid sequences from de novo mass spectrometry and Edman degradation with an expressed sequence tag library dataset of C. rubrum. Scleritin is a secreted basic phosphorylated protein which exhibits a short amino acid sequence of 135 amino acids and a signal peptide of 20 amino acids. From specific antibodies raised against peptide sequences of scleritin, we obtained immunolabeling of scleroblasts and OM of the sclerites which provides information on the biomineralization pathway in C. rubrum.
我们首次从柳珊瑚的有机基质(OM)中分离和鉴定了一种蛋白质。这种名为硬蛋白的蛋白质是从 OM 的 EDTA 可溶部分提取的主要蛋白质之一。通过比较从头测序的质谱和 Edman 降解的氨基酸序列与柳珊瑚的表达序列标签文库数据集,获得了完整的开放阅读框(ORF)。硬蛋白是一种分泌的碱性磷酸化蛋白,其氨基酸序列短,为 135 个氨基酸,信号肽为 20 个氨基酸。针对硬蛋白肽序列制备的特异性抗体进行免疫标记,我们获得了硬骨细胞和骨片 OM 的免疫标记,这为柳珊瑚的生物矿化途径提供了信息。