Objective: To analyze the amino acid residue sequences of adeno-associated virus capsid protein (VP1) of different serotypes, and establish the similarity comparison method and database, and to predict the serotypes that VP1 may belong to after amino acid residue sequence mutation. Methods: The amino acid residues of VP1 of adeno-associated virus of different serotypes were analyzed to find out a common sequence, and the sequences were aligned. The hydrophobic amino acid residues, hydrophilic amino acid residues and acid/base amino acid residues were assigned separately. The assignment parameters included side chain length, group number and dissociation/binding strength. Then the similarity was calculated by correlation coefficient method and difference method. The global charge was defined to determine the boundary value, and determine whether the change of charge will cause a significant change in the electric field around the protein. Finally, considering the relationship of the four, the similarity with the known serological sequences was given. The protein amino acid sequences of other known serotypes were verified. Results: Basically, different serotypes could be distinguished, and different subtypes of the same serotype have a certain tolerance. Conclusion: The relationship between the amino acid residues sequence of adeno-associated VP1 and its serotype was discussed, hoping to provide support for the research and development of recombinant adeno-associated virus as a vector for gene therapy.
HOU Zong-wen, QIN Xi, TAO Lei, SHI Xin-chang, ZHANG Yi-xuan
. Sequence analysis of amino acid residues in adeno-associated virus capsid protein of different serotypes*[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(12)
: 2017
-2024
.
DOI: 10.16155/j.0254-1793.2023.12.04
[1] 国家药品监督管理局药品审评中心. 体内基因治疗产品药学研究与评价技术指导原则(试行)[EB/OL]. (2022-5-26) [2023-06-14]. https://www.cde.org.cn/main/news/viewInfoCommon/c0ec5e347ba84df67bf75e15f6ad3f3f
Enter For Drug Evaluation. Technical Guidelines for Pharmaceutical Research and Evaluation of In Vivo Gene Therapy Products (Trial)[DB/OL]. (2022-5-26) [2023-06-14]. https://www.cde.org.cn/main/news/viewInfoCommon/c0ec5e347ba84df67bf75e15f6ad3f3f
[2] MIETZSCH M, HULLJA, MAKAL VE, et al. Characterization of the serpentine adeno-associated virus (SAAV) capsid structure: receptor interactions and antigenicity[J]. J Virol, 2022, 96(11):e00335
[3] GILES AR, SIMS JJ, TURNER KB, et al. Deamidation of amino acids on the surface of adeno-associated virus capsids leads to charge heterogeneity and altered vector function[J]. Mol Ther, 2018, 26(12):2848
[4] LARGE EE, SILVERIA MA, ZANE GM, et al. Adeno-associated virus (AAV) gene delivery: dissecting molecular interactions upon cell entry[J]. Viruses, 2021, 13(7):1336
[5] HERRMANN AK, GRIMM D. High-throughput dissection of AAV-host interactions: the fast and the curious[J]. Mol Biol, 2018, 430(17):2626
[6] 曹雪涛.医学免疫学[M]. 第6版.北京:人民卫生出版社, 2013: 35
CAO XT, Medical Immunology[M]. 6nd Ed.Beijing:People’s Medical Publishing House, 2013:35
[7] 李响, 史新昌, 范文红, 等. 重组人促红素异构体唾液酸化程度分析[J]. 中国药学杂志, 2018, 53(19):1682
LI X, SHI XC, FAN WH, et al. Estimation of the degree of salivary acidification of recombinant-human-erythropoietin (rhEPO) [J]. Chin Pharm J, 2018, 53(19):1682
[8] 史新昌, 李响, 范文红, 等. 重组白介素-15融合蛋白成像毛细管等电聚焦电泳谱图分析[J]. 中国药学杂志, 2020(9):745
SHI XC, LI X, YU L, et al. Analysis of WCID-cIEF spectrogram of recombinant interleukin-15 fusion protein[J]. Chin Pharm J, 2020 (9):745
[9] 汤其群.生物化学与分子生物学[M]. 上海:复旦大学出版社, 2015:9
TANG QQ.Biochemistry and Molecular Biology[M]. Shanghai:Fudan University Press, 2015:9
[10] 丁明孝, 王喜忠, 张传茂, 等.细胞生物学[M]. 第5版.北京:高等教育出版社, 2020:5
DING MX, WANG XZ, ZHANG CM, et al. Cell Biology[M]. 5nd EG.Beijing:Higher Education Rress, 2020:5
[11] 史新昌, 李响, 于雷, 等. 重组人促红素等电点异构体分子体内活性强度分析[J]. 中国生物制品学杂志, 2020, 133(5):549
SHI XC, LI X, YU L, et al. In vivo biological activity strength of isoelectric point isoform molecules of recombinant human erythropoietin[J]. Chin J Biologicals, 2020, 33(5):549
[12] 林训忠.酸碱度与人体健康[J]. 科技信息, 2013(2):16
LIN XZ.pH and human health[J]. Scientific and technical information, 2013(2):16
[13] MINDELL JA. Lysosomal acidification mechanisms[J]. Annu Rev Physiol, 2012, 74:69
[14] CANTON J, GRINSTEIN S. Measuring lysosomal pH by fluorescence microscopy[J]. Methods Cell Biol, 2015;126:85
[15] BRORSON K, KREJCI S, LEE K, et al. Bracketed generic inactivation of rodent retroviruses by low pH treatment for monoclonal antibodies and recombinant proteins[J]. Biotechnol Bioengin, 2008, 32(3):321
[16] LIU LK, BECKER DF, TANNER JJ. Structure, function, and mechanism of proline utilization A (PutA)[J]. Arch Biochem Biophys, 2017, 632:142
