基于PacBio三代测序技术鉴定cisAB亚型及利用分子对接探究其分子机制

doi:10.3969/j.issn.1671-2587.2024.06.014

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (6): 792-796.DOI: 10.3969/j.issn.1671-2587.2024.06.014

基于PacBio三代测序技术鉴定cisAB亚型及利用分子对接探究其分子机制

夏悦昕¹, 邵林楠¹, 李宁¹, 周世航¹, 刘志远²

¹大连市血液中心;
²大连市妇女儿童医疗中心（集团）儿童医院院区,辽宁大连 116000

收稿日期:2024-05-21 出版日期:2024-12-20 发布日期:2024-12-20
通讯作者: 周世航,主要从事免疫血液学研究工作,（E-mail）zshsail@163.com。刘志远,主要从事二代测序研究,（E-mail）15842622503@126.com。
作者简介:夏悦昕,主要从事免疫血液学研究工作,（E-mail）xiayuexin001@163.com。

Identify the CisAB Subtype by PacBio Third-generation Sequencing Technology and Explore the Molecular Mechanism by Molecular Docking

XIA Yuexin¹, SHAO Linnan¹, LI Ning¹, ZHOU Shihang¹, LIU zhiyuan²

¹Dalian Blood Center, Dalian 116000,
²Dalian Municipal Women and Children's Medical Center (Group), Dalian 116000

Received:2024-05-21 Online:2024-12-20 Published:2024-12-20

摘要/Abstract

摘要： 目的利用血清学及PacBio三代测序技术鉴定1例罕见cisAB亚型,利用分子对接探讨糖基转移酶氨基酸突变导致亚型的分子机制。方法利用免疫血清学试验鉴定ABO表型,利用PacBio三代测序方法鉴定其基因型。突变后的糖基转移酶同源建模和分子对接后,与野生型α1,3-N-乙酰氨基半乳糖转移酶（α1-3-N-acetylgalactosaminyltransferase,GTA）进行结构叠加和比对,探究关键位点氨基酸突变后糖基转移酶的结构变化。结果免疫血清学试验鉴定表型为AB型,三代基因测序技术结果显示该患者基因型为cisAB.01/O.01.02,分子对接结果显示与野生型相比,cisAB01糖基转移酶p.Pro156Leu和p.Gly268Ala突变导致临近氨基酸结构朝向改变,催化口袋凹槽结构改变,出现双功能酶活性。结论 p.Pro156Leu和p.Gly268Ala突变是形成cisAB01糖基转移酶双功能活性的关键位点。

关键词: cisAB亚型, PacBio三代测序, 同源建模, 分子机制, 分子对接

Abstract: Objective To identify a rare cisAB subtype by immunoserology and PacBio third-generation sequencing technology, and explore the molecular mechanism of amino acid mutations in cisAB glycosyltransferase by molecular docking. Methods The ABO phenotype was identified by immunoserological method, and the genotype was identified by three generation sequencing technology. The homologous modeling and molecular docking of cisAB01 glycosyltransferase were used to explore the structural changes of the glycosyltransferase after key amino acid mutations. Results The serological phenotype was AB. The gene sequencing showed that the genotype was cisAB.01/O.01.02. Homology modeling and molecular docking showed that p.Pro156Leu and p.Gly268Ala mutation of cisAB01 glycosyltransferase alter the spatial structure of the binding groove of catalytic active center. Conclusion The p.Pro156Leu and p.Gly268Ala mutation are key sites for the bifunctional activity of cisAB01 glycosyltransferase．

Key words: cisAB subtype, PacBio third-generation sequencing technology, Homology modeling, Molecular mechanism, Molecular docking

中图分类号:

R331.1⁺41

夏悦昕, 邵林楠, 李宁, 周世航, 刘志远. 基于PacBio三代测序技术鉴定cisAB亚型及利用分子对接探究其分子机制[J]. 临床输血与检验, 2024, 26(6): 792-796.

XIA Yuexin, SHAO Linnan, LI Ning, ZHOU Shihang, LIU zhiyuan. Identify the CisAB Subtype by PacBio Third-generation Sequencing Technology and Explore the Molecular Mechanism by Molecular Docking[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(6): 792-796.

参考文献

[1] 丁洁岚,冯娟,陈皆锋,等. 产科大量输血方案的临床研究进展[J]. 中华全科医学,2022,20(3):468-472.
[2] 崔文燕,刘金华,付威义. 利用三代测序技术鉴定c.586T>C突变导致的Bel亚型[J]. 临床输血与检验,2024,26(2):262-266.
[3] SHAO L N,XIA Y X,YANG Y C,et al.PacBio third-generation sequencing reveals an ABO gene promoter mutation,c.-35_-18del,leading to weakened B antigen expression[J]. Ann Lab Med,2024.
[4] SHAO L N,YANG Y C,XIA Y X,et al.Novel missense mutation c.797T>C (p.Met266Thr) gives rise to the rare B(A) phenotype in a Chinese family[J]. Vox Sang,2024,119(4):383-387.
[5] 朱永亮,李莺,许进明,等. 血清学结合分子生物学方法鉴定ABO*cisAB.01/O.01.01血型1例[J]. 临床血液学杂志,2020,33(12):863-866.
[6] 赵倩,苏蔓,李茵,等. B(A)血型与cisAB血型的血清学表型及基因型研究[J]. 临床血液学杂志,2020,33(8):521-524.
[7] 马晓莉,耿明璐,李建斌,等. 12例CisAB亚型表型与基因分型分析[J]. 中国输血杂志,2022,35(12):1204-1207.
[8] WANG C B,TANG Y C,ZHANG P P,et al.Detection and phenotype analysis of a novel Ael blood group allele[J]. Vox Sang,2024,119(1):74-78.
[9] PATENAUDE S I,SETO N O L,BORISOVA S N,et al. The structural basis for specificity in human ABO(H) blood group biosynthesis[J]. Nat Struct Biol,2002,9:685-690.
[10] 孔永奎,杨乾坤,王书亚,等. cisAB03亚型分子生物学鉴定及分子对接[J]. 郑州大学学报(医学版),2020,55(4):533-536.
[11] MARTINEZ X,KRONE M,ALHARBI N,et al.Molecular graphics:bridging structural biologists and computer scientists[J]. Structure,2019,27(11):1617-1623.
[12] CAI X H,JIN S,LIU X,et al.Molecular genetic analysis for the B subgroup revealing two novel alleles in the ABO gene[J]. Transfusion,2008,48(11):2442-2447.
[13] Daniels G.Human blood groups[M]. 2nd ed. Oxford: Blackwell Science,2002.
[14] 杨乾坤,靳慧芳,吕先萍,等. CisAB血型的血清学鉴定及分子遗传学研究: 附1例报告[J]. 中国输血杂志,2019,32(1):38-41.
[15] YAMAMOTO F,MCNEILL P D,HAKOMORI S.Human histo-blood group A2 transferase coded by A2 allele,one of the A subtypes,is characterized by a single base deletion in the coding sequence,which results in an additional domain at the carboxyl terminal[J]. Biochem Biophys Res Commun,1992,187(1):366-374.

基于PacBio三代测序技术鉴定cisAB亚型及利用分子对接探究其分子机制

Identify the CisAB Subtype by PacBio Third-generation Sequencing Technology and Explore the Molecular Mechanism by Molecular Docking

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