Bicocchi Maria P, Pasino Mirella, Lanza Tiziana, Bottini Federico, Boeri Elio, Mori Pier G, Molinari Angelo C, Rosano Camillo, Acquila Maura
Haemostasis and Haemophilia Laboratory, IV Paediatric Department, G. Gaslini Institute, Genova, Italy.
Br J Haematol. 2003 Sep;122(5):810-7. doi: 10.1046/j.1365-2141.2003.04494.x.
We describe 18 novel mutations, unreported in the Haemophilia A mutation Databases, that have been identified in a cohort of unrelated, Italian patients affected with haemophilia A (HA). Screening of the factor VIII gene (FVIII) was performed using denaturing high-performance liquid chromatography (DHPLC) and direct sequencing. Eight mutations were characterized as non-missense alterations, and the remaining 10 were missense mutations. Heterozygosity for the identified mutations was observed in the female relatives of patients belonging to eight families with sporadic cases. In an attempt to understand better the causative effect of the mutations and the clinical variability of the patients, missense mutation consequences were investigated for: (1) the nature of the new amino acid; (2) the location of the substituted amino acid within crystallographic and theoretical models; and (3) the degree of conservation of the native residue in factor VIII (FVIII) protein and FVIII-related protein family aligned sequences. These research tools have provided evidence that the mutations we describe involve residues that were conserved, at least in FVIII proteins, in all the species we compared.
我们描述了18种在血友病A突变数据库中未报告的新突变，这些突变是在一组患血友病A（HA）的不相关意大利患者中鉴定出来的。使用变性高效液相色谱（DHPLC）和直接测序对凝血因子VIII基因（FVIII）进行筛查。8种突变被鉴定为无义改变，其余10种为错义突变。在8个散发病例家庭的患者女性亲属中观察到所鉴定突变的杂合性。为了更好地理解这些突变的致病作用以及患者的临床变异性，我们研究了错义突变的后果：（1）新氨基酸的性质；（2）取代氨基酸在晶体学和理论模型中的位置；（3）凝血因子VIII（FVIII）蛋白和FVIII相关蛋白家族比对序列中天然残基的保守程度。这些研究工具提供了证据，表明我们描述的突变涉及至少在FVIII蛋白中，在我们比较的所有物种中都保守的残基。
