高通量质谱法用于小鼠器官内源性肽的鉴定
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摘要
内源性肽以细胞因子、生长激素、激素肽等形式在人体的内分泌、神经、细胞生长和生殖各个领域发挥功能。神经肽是一种内源性肽,与痛觉、睡眠、情绪、学习与记忆等生理活动相关,不但存在于脑部神经细胞,也存在于其他体液和器官内并发挥重要作用。目前对器官内源性肽的研究仍不足,尤其是其中的神经肽。文中应用液质联用串联质谱高通量鉴定胰腺、心脏、肝脏和肾脏中内源性肽的分布以及神经肽的种类。鉴定结果显示,在肝脏中内源性肽和神经肽的数目最多,而胰腺中最少;所鉴定到的内源性肽具有器官特异性,在4个器官中分别呈现不同的动态分布;4个器官中神经肽的LPV(最长肽变异体)数目差异较大,而且基因家族的分布也各不相同,比如胰腺中的神经肽多属于Glucagon家族,心脏中的神经肽分别属于ACBD7、Granins、PEBP等几个家族。鉴定结果将为疾病的机制研究和治疗药物的研发提供参考。
Endogenous peptides, in the form of cytokines, growth hormones and hormone peptides, play an important role in human hormones, nerves, cell growth and reproduction. Neuropeptide is a kind of endogenous peptide, which is related to the physiological activities of pain, sleep, emotion, learning and memory. Neuropeptides exist not only in the nerve cells of the brain, but also in other body fluids and organs. At present, there is still a lack of research on endogenous peptides, especially on neuropeptides.In this study, high-throughput liquid chromatography tandem mass spectrometry was used to identify the distribution of endogenous peptides in the pancreas, heart, liver and kidney as well as the types of neuropeptides. The results showed that the number of endogenous peptides and neuropeptides in the liver was the highest while that of the pancreas was the lowest. The identified endogenous peptides were organ-specific and presented different dynamic distribution in four kinds of organs. The number of LPV(Longest peptide variant) of neuropeptide in the four organs varies greatly, and the distribution of gene family is also different. For example, neuropeptide in pancreas belongs to Glucagon family, while neuropeptide in heart belongs to ACBD7, Granins, PEBP and other families. The identification results will provide reference value for the mechanism study of diseases and the research and development of therapeutic drugs.
Endogenous peptides, in the form of cytokines, growth hormones and hormone peptides, play an important role in human hormones, nerves, cell growth and reproduction. Neuropeptide is a kind of endogenous peptide, which is related to the physiological activities of pain, sleep, emotion, learning and memory. Neuropeptides exist not only in the nerve cells of the brain, but also in other body fluids and organs. At present, there is still a lack of research on endogenous peptides, especially on neuropeptides.In this study, high-throughput liquid chromatography tandem mass spectrometry was used to identify the distribution of endogenous peptides in the pancreas, heart, liver and kidney as well as the types of neuropeptides. The results showed that the number of endogenous peptides and neuropeptides in the liver was the highest while that of the pancreas was the lowest. The identified endogenous peptides were organ-specific and presented different dynamic distribution in four kinds of organs. The number of LPV(Longest peptide variant) of neuropeptide in the four organs varies greatly, and the distribution of gene family is also different. For example, neuropeptide in pancreas belongs to Glucagon family, while neuropeptide in heart belongs to ACBD7, Granins, PEBP and other families. The identification results will provide reference value for the mechanism study of diseases and the research and development of therapeutic drugs.
引文
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[26]Sajdyk TJ,Shekhar A,Gehlert DR.Interactions between NPY and CRF in the amygdala to regulate emotionality.Neuropeptides,2004,38(4):225-234.
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[28]Camargo ACM,Fernandes BL,Cruz L,et al.Bioactive Peptides Produced by Limited Proteolysis.New York:Colloquium Series on Neuropeptides,2012.
[29]Burbach JPH.Neuropeptides from concept to online database www.neuropeptides.nl.Eur J Pharmacol,2010,626(1):27-48.
[30]Perry T,Lahiri DK,Sambamurti K,et al.Glucagon-like peptide-1 decreases endogenous amyloid-βpeptide(Aβ)levels and protects hippocampal neurons from death induced by Aβand iron.J Neurosci Res,2003,72(5):603-612.
[31]Della Valle B,Brix GS,Brock B,et al.Glucagon-like peptide-1 analog,liraglutide,delays onset of experimental autoimmune encephalitis in Lewis rats.Front Pharmacol,2016,7:433.
[2]Fricker LD.Neuropeptides and Other Bioactive Peptides:From Discovery to Function.New York:Morgan&Claypool Life Sciences,2012.
[3]H?kfelt T,Bartfai T,Bloom F.Neuropeptides:opportunities for drug discovery.Lancet Neurol,2003,2(8):463-472.
[4]MathèAA,Stenfors C,Brodin E,et al.Neuropeptides in brain:effects of microwave irradiation and decapitation.Life Sci,1990,46(4):287-293.
[5]Kormos V,Gaszner B.Role of neuropeptides in anxiety,stress,and depression:from animals to humans.Neuropeptides,2013,47(6):401-419.
[6]Mooney C,Haslam NJ,Pollastri G,et al.Towards the improved discovery and design of functional peptides:common features of diverse classes permit generalized prediction of bioactivity.PLoS ONE,2012,7(10):e45012.
[7]Sk?ld K,Svensson M,Kaplan A,et al.A neuroproteomic approach to targeting neuropeptides in the brain.Proteomics,2002,2(4):447-454.
[8]F?lth M,Sk?ld K,Svensson M,et al.Neuropeptidomics strategies for specific and sensitive identification of endogenous peptides.Mol Cell Proteomics,2007,6(7):1188-1197.
[9]Secher A,Kelstrup CD,Conde-Frieboes KW,et al.Analytic framework for peptidomics applied to large-scale neuropeptide identification.Nat Commun,2016,7:11436.
[10]Shao XF,Ma M,Chen RB,et al.Identification of mouse brain neuropeptides by high throughput mass spectrometry.Chin J Biotech,2018,34(4):594-601(in Chinese).邵先锋,马敏,陈瑞冰,等.高通量质谱法用于小鼠脑部神经肽的鉴定.生物工程学报,2018,34(4):594-601.
[11]Altelaar AFM,Mohammed S,Brans MAD,et al.Improved identification of endogenous peptides from murine nervous tissue by multiplexed peptide extraction methods and multiplexed mass spectrometric analysis.JProteome Res,2009,8(2):870-876.
[12]An ZM,Chen YD,Koomen JM,et al.A mass spectrometry-based method to screen forα-amidated peptides.Proteomics,2012,12(2):173-182.
[13]Schmidlin T,Boender AJ,Frese CK,et al.Diet-induced neuropeptide expression:feasibility of quantifying extended and highly charged endogenous peptide sequences by selected reaction monitoring.Anal Chem,2015,87(19):9966-9973.
[14]Che FY,Lim J,Pan H,et al.Quantitative neuropeptidomics of microwave-irradiated mouse brain and pituitary.Mol Cell Proteomics,2005,4(9):1391-1405.
[15]Buchberger A,Yu Q,Li LJ.Advances in mass spectrometric tools for probing neuropeptides.Ann Rev Anal Chem,2015,8(1):485-509.
[16]Yamaguchi H,Sasaki K,Satomi Y,et al.Peptidomic identification and biological validation of neuroendocrine regulatory peptide-1 and-2.J Biol Chem,2007,282(36):26354-26360.
[17]van Dijck A,Hayakawa E,Landuyt B,et al.Comparison of extraction methods for peptidomics analysis of mouse brain tissue.J Neurosci Methods,2011,197(2):231-237.
[18]F?lth M,Sk?ld K,Norrman M,et al.SwePep,a database designed for endogenous peptides and mass spectrometry.Mol Cell Proteomics,2006,5(6):998-1005.
[19]Kim Y,Bark S,Hook V,et al.NeuroPedia:neuropeptide database and spectral library.Bioinformatics,2011,27(19):2772-2773.
[20]Wang Y,Wang MX,Yin SW,et al.NeuroPep:a comprehensive resource of neuropeptides.Database,2015,2015:bav038.
[21]Wardman JH,Berezniuk I,Di S,et al.ProSAAS-derived peptides are colocalized with neuropeptide Y and function as neuropeptides in the regulation of food intake.PLo SONE,2011,6(12):e28152.
[22]Banerjee I,Zhang JL,Moore-Morris T,et al.Thymosin beta 4 is dispensable for murine cardiac development and function.Circulat Res,2012,110(3):456-464.
[23]Marcos-Contreras OA,de Lizarrondo SM,Bardou I,et al.Hyperfibrinolysis increases blood-brain barrier permeability by a plasmin-and bradykinin-dependent mechanism.Blood,2017,128(20):2423-2434.
[24]Wang XH,Li L,H?lscher C,et al.Val8-glucagon-like peptide-1 protects against Aβ1-40-induced impairment of hippocampal late-phase long-term potentiation and spatial learning in rats.Neuroscience,2010,170(4):1239-1248.
[25]Broberger C,Johansen J,Johansson C,et al.The neuropeptide Y/agouti gene-related protein(AGRP)brain circuitry in normal,anorectic,and monosodium glutamate-treated mice.Proc Natl Acad Sci U S A,1998,95(25):15043-15048.
[26]Sajdyk TJ,Shekhar A,Gehlert DR.Interactions between NPY and CRF in the amygdala to regulate emotionality.Neuropeptides,2004,38(4):225-234.
[27]Tatemoto K,Carlquist M,Mutt V.Neuropeptide Y-Anovel brain peptide with structural similarities to peptide YY and pancreatic polypeptide.Nature,1982,296(5858):659-660.
[28]Camargo ACM,Fernandes BL,Cruz L,et al.Bioactive Peptides Produced by Limited Proteolysis.New York:Colloquium Series on Neuropeptides,2012.
[29]Burbach JPH.Neuropeptides from concept to online database www.neuropeptides.nl.Eur J Pharmacol,2010,626(1):27-48.
[30]Perry T,Lahiri DK,Sambamurti K,et al.Glucagon-like peptide-1 decreases endogenous amyloid-βpeptide(Aβ)levels and protects hippocampal neurons from death induced by Aβand iron.J Neurosci Res,2003,72(5):603-612.
[31]Della Valle B,Brix GS,Brock B,et al.Glucagon-like peptide-1 analog,liraglutide,delays onset of experimental autoimmune encephalitis in Lewis rats.Front Pharmacol,2016,7:433.