驴血清α1酸性糖蛋白生物信息学分析与纯化鉴定研究

doi:10.16431/j.cnki.1671-7236.2021.08.003

Abstract

Abstract: This study was aimed to carry out the bioinformatics prediction and analysis of alpha-1-acid glycoprotein(α1-AGP) of donkey, and an efficient purification method of α1-AGP from donkey was established.Analysis of donkey by bioinformatics online software.The basic physical, chemical properties and the secondary and tertiary structures of α1-AGP were predicted.According to the results of the analysis, the method of cation chromatography gel filtration chromatography was used to purify donkey α1-AGP, after SDS-PAGE, matrix assisted laser desorption ionization-time offlight mass spectrometry (MALDI-TOF-MS) was used to identify donkey α1-AGP.The results showed that donkey α1-AGP was stable and hydrophilic, composed of 201 amino acids, the theoretical isoelectric point(pI) was 4.99, there were 5 potential N-glycosylation sites, and the average value of hydrophilicity(GRAVY) was -0.386.α-helix and β-sheet appeared orderly in the secondary structure, accounting for 88.56%, the GMQE value and the Z value of QMEAN were used to verify that the model was in a reasonable range.After purification by cation exchange chromatography and gel filtration chromatography, a total of 50 mL protein solution was obtained, and a clear protein strip was obtained after SDS-PAGE.After MALDI-TOF-MS and retrieved from NCBI database, the protein with the highest score was donkey α1-AGP, which was much higher than other proteins, indicating that α1-AGP was the main component in the purified protein.In this study, we proposed a new method for the purification and identification of α1-AGP from donkeys, which had the advantages of high purification rate, simple operation and less samples needed, and laid a theoretical foundation for the subsequent biological research and application of α1-AGP from donkeys.

Key words: donkey; alpha-1-acid glycoprotein (α1-AGP); bioinformatics; purification

CLC Number:

S822

FEI Yaqi, TANG Chi, DENG Liang. Bioinformatics Analysis,Purification and Identification of Alpha-1-acid Glycoprotein in Serum of Donkey[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2705-2712.

References

[1] 李杰,王玉斌.中国驴业发展现状、问题及对策[J].中国畜牧杂志,2019,55(5):159-162. LI J,WANG Y B.Current situation,problems and countermeasures of donkey industry in China[J].Chinese Journal of Animal Science,2019,55(5):159-162.(in Chinese)
[2] FOURNIER T,MEDJOUBI-N N,PORQUET D.Alpha-1-acid glycoprotein[J].Biochimica et Biophysica Acta-Proteins and Proteomics,1990,1482(1-2):157-171.
[3] MACKIEWICZ A,MACKIEWICZ K.Glycoforms of serum α1-acid glycoprotein as markers of inflammation and cancer[J].Glycoconjugate Journal,1995,12(3):241.
[4] KRAUER B,DAYER P,ANNER R.Changes in serum albumin and α1-acid glycoprotein concentrations during pregnancy:An analysis of fetal-maternal pairs[J].BJOG,1984,91(9):875-881.
[5] GOLUB M S,KAAEKUAHIWI M A.Changes in plasma alpha 1-acid glycoprotein and albumin concentrations during late pregnancy in rhesus monkeys[J].International Journal of Clinical Chemistry,1997,262(1-2):29-37.
[6] HONDA M,OMORI Y,MINEI S,et al.Quantitative analysis of serum α1-acid glycoprotein levels in normal and diabetic pregnancy[J].Diabetes Research and Clinical Practice,1990,10(2):147-152.
[7] SMITH S A,WATERS N J.Pharmacokinetic and pharmacodynamic considerations for drugs binding to alpha-1-acid glycoprotein[J].Archives of Pharmacal Research,2018,36(2):30.
[8] RIKIHISA Y,YAMAMOTO S,KWAK I,et al.C-reactive protein and α1-acid glycoprotein levels in dogs infected with Ehrlichia canis[J].Journal of Clinical Microbiology,1994,32(4):912-917.
[9] PEEBLES E D,JACOB R,BRANTON S L,et al.Effects of mycoplasma gallisepticum vaccination on serum α1-acid glycoprotein concentrations in commercial layer chickens[J].Poultry Science,2014,93(6):1396-1402.
[10] RINALDI M,CECILIANI F,LECCHI C,et al.Differential effects of alpha1-acid glycoproteinon bovine neutrophil respiratory burst activity and IL-8 production[J].Veterinary Immunology & Immuno-pathology,2008,126(3-4):199-210.
[11] CAPERNA T J,SHANNON A E,BLOMBERG L A,et al.Identification of alpha-1 acid glycoprotein (AGP) as a potential marker of impaired growth in the newborn piglet[J].Reproduction Fertility and Development,2013,25(8):1126-1133.
[12] WATANABE M,IWAI K,SHIBATA S,et al.Purification and characterization of mouse alpha 1-acid glycoprotein and its possible role in the antitumor activity of some lichen polysaccharides[J].Chemical & Pharmaceutical Bulletin,1986,34(6):2532-2541.
[13] CECILIANI F,LECCHI C.The immune functions of α1-acid glycoprotein[J].Current Protein and Peptide Science,2019,20(6):505-524.
[14] GAMAGE D G,GUNARATNE A,PERIYANNAN G R,et al.Applicability of instability index for in vitro protein stability prediction[J].Protein and Peptide Letters,2019,26(5):339-347.
[15] JAYAPRAKASH N G,SUROLIA A.Role of glycosylation in nucleating protein folding and stability[J].Biochemical Journal,2017,474(14):2333-2347.
[16] LAURENT P,MIRIBEL L,BIENVENU J,et al.A three-step purification of human α1-acid glycoprotein[J].FEBS Letters,1984,168(1):79-83.
[17] SUCCARI M,MARIE-JOSÉ F,PERCHERON F.Microheterogeneity of alpha 1-acid glycoprotein:Variation during the menstrual cycle in healthy women,and profile in women receiving estrogen-progestogen treatment[J].Clinica Chimica Acta,1990,187(3):235-241.
[18] HEDBERG Y S,DOBRYDEN I,CHAUDHARY H,et al.Synergistic effects of metal-induced aggregation of human serum albumin[J].Colloids and Surfaces B-Biointerfaces,2019,173:751-758.
[19] 李欣蔚,廖佳,董秀瑜,等.功能性肽的分离及富集研究进展[J].食品科学,2020,41(1):267-276. LI X W,LIAO J,DONG X Y,et al.A review of the separation and enrichment of functional peptides[J].Food Science,2020,41(1):267-276.(in Chinese)
[20] COSKUN O.Separation techniques:Chromatography[J].Northern Clinics of Istanbul,2016,3(2):156-160.
[21] RYAN D J,SPRAGGINS J M,CAPRIOLI R M.Protein identification strategies in MALDI imaging mass spectrometry:A brief review[J].Current Opinion in Chemical Biology,2019,48:64-72.
[22] 王磊,张然然,刘华淼,等.马鹿鹿茸不同部位差异蛋白质组学分析[J].畜牧兽医学报,2017,48(8):1401-1415. WANG L,ZHANG R R,LIU H M,et al.Comparative proteomics analysis on different parts of cervus elaphus songaricus velvet antler[J].Chinese Journal of Animal and Veterinary Sciences,2017,48(8):1401-1415.(in Chinese)
[23] MATUOZZO M,SPAGNUOLO M S,HUSSEIN H A,et al.Novel biomarkers of mastitis in goat milk revealed by MALDI-TOF-MS-based peptide profiling[J].Biology-Basel,2020,9(8):193.
[24] HYKOLLARI A,MALZL D,WILSON I B H,et al.Protein-specific analysis of invertebrate glycoproteins[J].Methods in Molecular Biology,2019,1871:421-435.

Bioinformatics Analysis,Purification and Identification of Alpha-1-acid Glycoprotein in Serum of Donkey

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