Structural and functional characterization of novel F7 mutations identified in Chinese factor VII‐deficient patients
Structural and functional characterization of novel F7 mutations identified in Chinese factor VII‐deficient patients
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DOI：
10.1111/bjh.18768
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发表时间：
2023-03
影响因子：
6.5
通讯作者：
Can Lou;Jiali Jiang;Wei-Zhi Chen;Zhili Zhang;Guan-qun Xu;Yu Liu;J. Dai;Q. Ding;Xuefeng Wang;H. Wei;Youwei Wu;Qinggang Xu;Wenman Wu
中科院分区：
文献类型：
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作者：
Can Lou;Jiali Jiang;Wei-Zhi Chen;Zhili Zhang;Guan-qun Xu;Yu Liu;J. Dai;Q. Ding;Xuefeng Wang;H. Wei;Youwei Wu;Qinggang Xu;Wenman Wu
遗传性因子VII（FVII）缺乏症是一种罕见的隐性出血性疾病，估计患病率为1/500 000。我们调查了50例无血缘关系的中国FVII缺乏症患者，共发现25个F7基因突变，包括18个错义突变和5个剪接突变。外显子9的核苷酸转换c.1224T>G（p.His408Gln）构成突变热点，19例患者具有这种遗传变异。少数患者为有害突变的纯合子或复合杂合子，如无义突变、大插入或缺失，表明FVII完全缺乏可能与生命不相容。通过体外表达和功能研究，进一步分析了本研究中发现的8个新突变，包括1个小缺失（p.Glu49GlyfsTer101），3个I型错义突变，p.Cys238Phe，p.Gly420Asp，p.Ala252Val和4个II型错义突变，p.Val336Met，p.Ser342Gly，p.Gly432Ser和p.Ile213Asn。实验室表型和结构分析证实了p.Ile213Asn突变涉及切割和激活位点的功能后果。分子动力学模拟和结合能计算沿着以及p.Gly432 Ser突变的功能探测揭示了Gly 432残基在活化的凝血因子VII（凝血因子VIIa）与组织因子结合中的关键作用。
Hereditary factor VII (FVII) deficiency is a rare recessive bleeding disorder with an estimated prevalence of 1/500 000. We had investigated 50 unrelated Chinese patients with FVII deficiency and identified, in total, 25 mutations, including 18 missense mutations and 5 splicing mutations, on the F7 gene. The nucleotide transition c.1224T>G (p.His408Gln) in exon 9 constitutes a hotspot of mutation, with 19 patients harbouring this genetic variance. Few patients were homozygous or compound heterozygous for deleterious mutations, such as non‐sense mutations, large insertion or deletions, indicating that complete deficiency of FVII may not be compatible with life. The eight novel mutations identified in the study, including one small deletion (p.Glu49GlyfsTer101), three type I missense mutations, p.Cys238Phe, p.Gly420Asp, p.Ala252Val and four type II missense mutations, p.Val336Met, p.Ser342Gly, p.Gly432Ser and p.Ile213Asn, were further analysed by in vitro expression and functional studies. The laboratory phenotype and structural analysis confirmed the functional consequence of p.Ile213Asn mutation involving cleavage and activation site. The molecular dynamic simulations and binding energy calculations along with functional probing of p.Gly432Ser mutation revealed the critical role of residue Gly432 in the binding between activated factor VII (factor VIIa) and tissue factor.