不同月龄新西兰兔肝中差异蛋白质鉴定与验证

doi:10.3969/j.issn.1004-1524.20221467

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (10): 2299-2310.DOI: 10.3969/j.issn.1004-1524.20221467

不同月龄新西兰兔肝中差异蛋白质鉴定与验证

刘方程^a^,^b(), 王峰^a^,^b, 毕冬琳^a^,^b, 杨东亮^a^,^b, 杨晓莉^a^,^b, 柏家林^a^,^b, 李琼毅^a^,^b^,^*()

a.西北民族大学生物医学研究中心,生物工程与技术国家民委重点实验室,甘肃兰州 730030
b.西北民族大学生命科学与工程学院,甘肃兰州 730030

收稿日期:2022-10-25 出版日期:2023-10-25 发布日期:2023-10-31
作者简介:刘方程(1997—),男,甘肃兰州人,硕士研究生,从事病毒天然免疫机制研究。E-mail:956636176@qq.com
通讯作者: ^*李琼毅,E-mail:lqy@xbmu.edu.cn
基金资助:
国家自然科学基金(31702234);国家自然科学基金(31860696);西北民族大学中央高校基本科研业务费项目(31920190003);甘肃省教育厅高等教育教学成果培育项目(2020GSJXCGPY-07);西北民族大学教学改革研究项目(2020YBJG-43)

Identification and validation of differential proteins in liver of New Zealand rabbits at different months of age

LIU Fangcheng^a^,^b(), WANG Feng^a^,^b, BI Donglin^a^,^b, YANG Dongliang^a^,^b, YANG Xiaoli^a^,^b, BAI Jialin^a^,^b, LI Qiongyi^a^,^b^,^*()

a. Biomedical Research Center, Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China
b. Biomedical Research Center, College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China

Received:2022-10-25 Online:2023-10-25 Published:2023-10-31

摘要/Abstract

摘要：

兔出血症是由兔出血症病毒(Rabbit haemorrhagic disease virus, RHDV)引起的一种兔急性、高度致死性传染病,成年兔死亡率超过90%,但2周龄幼兔不发病。基于幼年兔与成年兔对RHDV的易感性差异,采用串联质谱标记(tadem mass tags, TMT)蛋白质组学技术对幼年兔(2周龄)和成年兔(6月龄)肝中的蛋白质进行检测和定量分析,从幼年兔和成年兔肝中定量获得4 353个蛋白质,筛选出821个差异蛋白质;相较于幼年兔,成年兔肝中294个蛋白质显著上调,527个蛋白质显著下调。GO功能富集分析结果显示,生物学过程主要富集到145个小分子代谢蛋白质、111个与氧化还原过程相关的蛋白质和478个参与代谢过程的蛋白质,细胞组分主要富集到528个细胞外成分蛋白质、139个线粒体蛋白质和366个细胞质蛋白质等,分子功能主要富集到342个催化活性蛋白质、93个氧化还原活性蛋白质和50个辅酶结合蛋白质等。821个差异蛋白质在KEGG Pathway数据库中共注释到47条KEGG信号通路,主要涉及代谢途径、糖代谢、氧化磷酸化作用和剪切小体等通路。幼年兔和成年兔肝中差异蛋白质互作网络分析发现,MRPS15的关联度最高,且差异蛋白质胆固醇酰基转移酶1(ACAT1)在互作网络中具有更多相互作用关系。从蛋白质水平探讨幼年兔和成年兔的感染差异机制,筛选并分析幼年兔和成年兔肝中的差异蛋白质,筛选出的差异蛋白质有望成为RHDV体外扩增细胞系构建的突破点,为构建适宜RHDV体外扩增的细胞系提供新的研究思路。

关键词: 新西兰兔, TMT蛋白质组学, 兔出血症病毒(RHDV), 肝, 差异蛋白质

Abstract:

Rabbit hemorrhagic disease is an acute and highly fatal infectious disease caused by Rabbit hemorrhagic disease virus (RHDV). The mortality rate of adult rabbits is more than 90%, but 2-week-old young rabbits are not affected. Based on the differences in the susceptibility of young rabbits and adult rabbits to RHDV, tandem mass tage (TMT) proteomics technology was used to detect and quantitatively analyze the proteins in liver of young rabbits (2 weeks old) and adult rabbits (6 months old). 4 353 proteins were quantitatively obtained from the liver of young rabbits and adult rabbits, and 821 differential proteins were screened. Among them, 294 proteins were significantly up-regulated and 527 proteins were significantly down-regulated in the liver of adult rabbits compared with young rabbits. The results of GO functional enrichment analysis showed that 145 small molecule metabolic proteins, 111 redox-related proteins and 478 proteins involved in metabolic processes were mainly enriched in biological processes; 528 extracellular component proteins, 139 mitochondrial proteins and 366 cytoplasmic proteins were mainly enriched in cellular components; and 342 catalytic active proteins, 93 redox active proteins and 50 coenzyme binding proteins were mainly enriched in molecular functions. A total of 47 KEGG (Kyoto Encyclopedia of Genes and Genomes) signaling pathways were annotated in 821 liver differential proteins in the KEGG pathway database, mainly involving metabolic pathways, carbon metabolism, oxidative phosphorylation and splicing bodies. The analysis of differential protein interaction network in liver of young and adult rabbits showed that MRPS15 had the highest correlation degree, and the differential protein cholesterol acyltransferase 1 (ACAT1) had more interaction in the interaction network. To explore the mechanism of infection difference between young and adult rabbits from the protein level, and to screen and analyze the differential proteins in liver of young and adult rabbits, it is expected to become a breakthrough point in the construction of RHDV in vitro amplification cell lines, and provide new research ideas for the construction of cell lines suitable for RHDV in vitro amplification.

Key words: New Zealand rabbit, tandem mass tage (TMT) proteomics, rabbit hemorrhagic disease virus (RHDV), liver, differential proteins

中图分类号:

S855.3

刘方程, 王峰, 毕冬琳, 杨东亮, 杨晓莉, 柏家林, 李琼毅. 不同月龄新西兰兔肝中差异蛋白质鉴定与验证[J]. 浙江农业学报, 2023, 35(10): 2299-2310.

LIU Fangcheng, WANG Feng, BI Donglin, YANG Dongliang, YANG Xiaoli, BAI Jialin, LI Qiongyi. Identification and validation of differential proteins in liver of New Zealand rabbits at different months of age[J]. Acta Agriculturae Zhejiangensis, 2023, 35(10): 2299-2310.

图/表 6

参考文献 35

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蛋白质ID Protein ID	蛋白质名称 Protein name	表达情况 Expression
Q8MI17	醇脱氢酶Aldehyde dehydrogenase 1 family member A1	上调Up-regulation
G1SS18	角蛋白8Keratin 8	上调Up-regulation
G1SIJ2	胆固醇酰基转移酶1 Acetyl-CoA acetyltransferase 1	下调Down-regulation
G1U758	角蛋白18 Keratin 18	上调Up-regulation
G1TT06	组蛋白结合蛋白1 Selenium binding protein 1	上调Up-regulation
G1SRF7	热休克蛋白9 Heat shock protein family A member 9	下调Down-regulation
G1SKT4	ATP合成酶、H+转运、线粒体F1复合体、α亚单位1	下调Down-regulation
	ATP synthase, H+transporting, mitochondrial F1 complex, alpha subunit 1
A0A5F9CMG2	3-羟基-3-甲基戊二酰-CoA合成酶2 3-Hydroxy-3-methylglutaryl-CoA synthase 2	下调Down-regulation
G1SXX5	羟基酰基-CoA脱氢酶三功能多酶复合体亚基α	下调Down-regulation
	Hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha
G1SJS1	组蛋白2簇H2be Histone 2 cluster H2be	下调Down-regulation
A0A5F9C4D8	UDP葡萄糖焦磷酸化酶2 UDP-glucose pyrophosphorylase 2	上调Up-regulation
G1T1U7	亚甲基四氢叶酸脱氢酶	上调Up-regulation
	Methylenetetrahydrofolate dehydrogenase, cyclohydrolase and formyltetrahydrofolate synthetase 1
U3KMH9	苹果酸脱氢酶2 Malate dehydrogenase 2	下调Down-regulation
P21195	脯氨酰4-羟化酶亚基beta Prolyl 4-hydroxylase subunit beta	下调Down-regulation
O18750	热休克蛋白90 beta家族成员1 Heat shock protein 90 beta family member 1	下调Down-regulation
G1T2A7	谷胱甘肽S转移酶α1 Glutathione S-transferase alpha I	上调Up-regulation
G1T295	环氧化物水解酶1 Epoxide hydrolase 1	上调Up-regulation
G1U723	前列腺素-E(2) 9还原酶样Prostaglandin-E(2) 9-reductase-like	上调Up-regulation
A0A5F9C5G3	丝氨酸羟甲基转移酶1 Serine hydroxymethyltransferase 1	上调Up-regulation
G1SNJ7	丙氨酸-乙醛酸氨基转移酶2 Alanine-glyoxylate aminotransferase 2	下调Down-regulation

蛋白质ID Protein ID	蛋白质名称 Protein name	表达情况 Expression
Q8MI17	醇脱氢酶Aldehyde dehydrogenase 1 family member A1	上调Up-regulation
G1SS18	角蛋白8Keratin 8	上调Up-regulation
G1SIJ2	胆固醇酰基转移酶1 Acetyl-CoA acetyltransferase 1	下调Down-regulation
G1U758	角蛋白18 Keratin 18	上调Up-regulation
G1TT06	组蛋白结合蛋白1 Selenium binding protein 1	上调Up-regulation
G1SRF7	热休克蛋白9 Heat shock protein family A member 9	下调Down-regulation
G1SKT4	ATP合成酶、H+转运、线粒体F1复合体、α亚单位1	下调Down-regulation
	ATP synthase, H+transporting, mitochondrial F1 complex, alpha subunit 1
A0A5F9CMG2	3-羟基-3-甲基戊二酰-CoA合成酶2 3-Hydroxy-3-methylglutaryl-CoA synthase 2	下调Down-regulation
G1SXX5	羟基酰基-CoA脱氢酶三功能多酶复合体亚基α	下调Down-regulation
	Hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha
G1SJS1	组蛋白2簇H2be Histone 2 cluster H2be	下调Down-regulation
A0A5F9C4D8	UDP葡萄糖焦磷酸化酶2 UDP-glucose pyrophosphorylase 2	上调Up-regulation
G1T1U7	亚甲基四氢叶酸脱氢酶	上调Up-regulation
	Methylenetetrahydrofolate dehydrogenase, cyclohydrolase and formyltetrahydrofolate synthetase 1
U3KMH9	苹果酸脱氢酶2 Malate dehydrogenase 2	下调Down-regulation
P21195	脯氨酰4-羟化酶亚基beta Prolyl 4-hydroxylase subunit beta	下调Down-regulation
O18750	热休克蛋白90 beta家族成员1 Heat shock protein 90 beta family member 1	下调Down-regulation
G1T2A7	谷胱甘肽S转移酶α1 Glutathione S-transferase alpha I	上调Up-regulation
G1T295	环氧化物水解酶1 Epoxide hydrolase 1	上调Up-regulation
G1U723	前列腺素-E(2) 9还原酶样Prostaglandin-E(2) 9-reductase-like	上调Up-regulation
A0A5F9C5G3	丝氨酸羟甲基转移酶1 Serine hydroxymethyltransferase 1	上调Up-regulation
G1SNJ7	丙氨酸-乙醛酸氨基转移酶2 Alanine-glyoxylate aminotransferase 2	下调Down-regulation

通路名称 Pathway name	ID	差异蛋白数量 Number of differential proteins
代谢途径Metabolic pathways	ocu01100	179
糖代谢Carbon metabolism	ocu01200	30
氧化磷酸化Oxidative phosphorylation	ocu00190	29
帕金森氏症Parkinson disease	ocu05012	29
剪切小体Spliceosome	ocu03040	26
阿兹海默病Alzheimer disease	ocu05010	30
丙酸代谢Propanoate metabolism	ocu00640	12
乙氧基化物和二甲酸酯代谢通路Glyoxylate and dicarboxylate metabolism	ocu00630	11
类固醇生物合成Steroid biosynthesis	ocu00100	8
丁酸盐代谢通路Butanoate metabolism	ocu00650	9

通路名称 Pathway name	ID	差异蛋白数量 Number of differential proteins
代谢途径Metabolic pathways	ocu01100	179
糖代谢Carbon metabolism	ocu01200	30
氧化磷酸化Oxidative phosphorylation	ocu00190	29
帕金森氏症Parkinson disease	ocu05012	29
剪切小体Spliceosome	ocu03040	26
阿兹海默病Alzheimer disease	ocu05010	30
丙酸代谢Propanoate metabolism	ocu00640	12
乙氧基化物和二甲酸酯代谢通路Glyoxylate and dicarboxylate metabolism	ocu00630	11
类固醇生物合成Steroid biosynthesis	ocu00100	8
丁酸盐代谢通路Butanoate metabolism	ocu00650	9

不同月龄新西兰兔肝中差异蛋白质鉴定与验证

Identification and validation of differential proteins in liver of New Zealand rabbits at different months of age

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