1.山西医科大学实验动物中心,太原 030001;2.山西医科大学基础医学研究中心,山西 晋中030600;3.山西医科大学附属心血管病医院心内科,太原 030024
R-33
1. Laboratory Animal Center, Shanxi Medical University, Taiyuan 030001, China. 2. School of Basic Medicine, Shanxi Medical University, Jinzhong 030600. 3. Department of Cardiology, Cardiovascular Hospital of Shanxi Medical University, Taiyuan 030024
目的 鉴定26S 蛋白酶体非ATP 酶调节亚基13(PSMD13)在Ⅱ型糖尿病中国地鼠的表达及对其理化性质、信号肽、亚细胞定位、功能结构域等进行分析,为深入研究其功能提供理论依据。 方法 荧光定量PCR 法检测PSMD13 mRNA 在中国地鼠和糖尿病地鼠肝、骨骼肌、脂肪和小肠组织的表达;PSMD13 氨基酸序列从NCBI 网站下载,氨基酸同源性分析采用DNAman 软件;PSMD13 蛋白的理化性质分析采用Prot Param 在线工具;PSMD13 蛋白的亲疏水性、信号肽、亚细胞定位采用Prot Scale 软件分析;PSMD13 蛋白结构域、二级结构分别采用Conserved Domain 数据库和SOPMA 工具分析;PSMD13 蛋白质的互作蛋白及功能网络采用STRING 数据库分析。 结果 RTqPCR结果显示PSMD13 mRNA 在糖尿病地鼠脂肪组织中表达下降;PSMD13 位于中国地鼠第3 号染色体,含有13个外显子,相对分子质量为42 942. 48,含有378 个氨基酸残基;PSMD13 理论等电点为6. 1,属酸性蛋白;其与小鼠、大鼠、果蝇、猩猩和人PSMD13 的氨基酸序列同源性高达97. 78%;PSMD13 蛋白质无信号肽、无跨膜结构,主要位于细胞质中;PSMD13 蛋白质羧基端含有一个PCI 结构域,与PSMD7、PSMD8、PSMA3、PSMC1 和USP14 等相互作用,参与泛素依赖性蛋白质分解代谢过程。 结论 PSMD13 在糖尿病中国地鼠脂肪组织中表达下降,该蛋白质在进化上高度保守,参与蛋白质的蛋白酶体降解途径,其表达下调可能参与糖尿病的发生发展。
Objective To identify mRNA expression of 26S proteasome non-ATPase regulatory subunit 13(PSMD13) in Chinese hamsters with type Ⅱ diabetes mellitus (T2DM), analyze its physicochemical properties, signal peptides, subcellular localization, and functional domains, and provide a theoretical basis to study its function. Methods PSMD13 mRNA expression in the liver, skeletal muscle, fat and small intestinal tissues of Chinese hamsters with or without T2DM was determined by RT-qPCR. The amino acid sequence of PSMD13 was downloaded from the NCBI website and amino acid homology analysis was performed using DNAman software. Physicochemical properties of PSMD13 were analyzed by the Prot Param online tool. Hydrophobicity, signal peptides, and subcellular localization of PSMD13 were analyzed by Prot Scale software. The functional domain and secondary structure of PSMD13 were analyzed using the Conserved Domain database and SOPMA tool, respectively. The interacting proteins and functional networks of PSMD13 were analyzed using STRING databases. Results RT-qPCR showed that PSMD13 mRNA expression was decreased in adipose tissue of T2MD Chinese hamsters compared with that in normal Chinese hamsters. PSMD13 was located on chromosome 3, contained 13 exons, encoded a protein containing 378 amino acid residues, and its relative molecular mass was 42 942. 48. PSMD13 was an acidic protein and its theoretical isoelectric point was 6. 1. Amino acid sequence homology of PSMD13 with mice, rats, Drosophilae melanogaster, orangutans, and humans was as high as 97. 78%. PSMD13 protein had no signal peptide or transmembrane structure, and was mainly located in the cytoplasm. The carboxyl terminal of the PSMD13 protein contained a PCI domain that interacted with PSMD7, PSMD8, PSMA3, PSMC1, and USP14 to participate in ubiquitin-dependent protein catabolism. Conclusions The PSMD13 protein is highly conserved in evolution and its expression is low in adipose tissue of T2DM Chinese hamsters. PSMD13 is involved in the proteasome degradation pathway, and its downregulation in adipose tissue may be involved in the occurrence and development of T2MD.
郭 民.中国地鼠26S 蛋白酶体非ATP 酶调节亚基13 的表达鉴定及生物信息学分析[J].中国比较医学杂志,2023,33(2):15~21.
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