马氏珠母贝磺基转移酶PmCHST11基因的分子特征与表达分析

Molecular Characterization and Expression Analysis of PmCHST11 Gene from Sulfotransferase of Pinctada martensii

硫酸软骨素(CS)具有抗炎、抗病毒和软骨保护功能等免疫学功能。硫酸软骨素磺基转移酶CHST11(carbohydrate sulfotransferase 11,chondroitin 4)催化磺酸基从供体PAPS(3'-磷酸腺苷酰-5'-磷酸硫酸盐)转移到软骨素N-乙酰半乳糖胺的4位碳上,实现磺酸化反应,是合成硫酸软骨素的关键酶。本研究通过对马氏珠母贝磺基转移酶基因Pm CHST11(Pinctada martensii carbohydrate sulfotransferase 11)的全长克隆及组织表达分析,探究Pm CHST11在马氏珠母贝免疫调节中的作用。结果表明:Pm CHST11序列全长为1 418 bp,编码426个氨基酸;Pm CHST11序列含有信号肽结构区域,跨膜结构域和磺基转移酶-2结构域,为高尔基体膜偶联磺基转移酶;多序列比对结果显示,Pm CHST11与其它物种的CHST11同源性较低;实时荧光定量结果表明,Pm CHST11基因在多个组织中均有表达,其中在外套膜边缘区显著高表达(p<0.05)。本研究为分析Pm CHST11在马氏珠母贝中的免疫调节中的作用积累基础资料。

Chondroitin sulfate （CS） has immune functions of anti-inflammatory, antiviral and cartilage protective. CHST11 （Carbohydrate sulfotransferase 11, chondroitin 4） catalyzes the sulfate transferation from a donor PAPs （3＇-adenosine phosphate-5＇-phosphate sulfate） to the position 4 of chondroitine repeating disaccharide unit N-acetyl galactose and accomplishes the sulfonation and it is the key enzyme in the synthesis of chondroitin sulfate. To explore the immune function of CHST11 gene in Pinctada martensii, the full-length sequence of PmCHST11 gene from sulfotransferase of Pinctada martensii was cloned and analyzed. The results showed that PmCHST11 sequence with full-length of 1 418 bp encoding 426 amino acids, and contained a signal peptide structure region, a transmembrane domain and a sulfotransferase-2 domain, which was the Golgi membrane coupled sulfonyl transferase. The results of multiple sequence alignments exhibited that PmCHSTll had low homology with CHST11 of other species. Real-time PCR showed that PmCHST11 gene was expressed in many tissues and was most highly expressed in the marginal zone of outer membrane （p〈0.05）. Above all, this study provides useful information for the analysis of the role of PmCHST11 in immune regulation of Pinctada martensii.