Loop-Sequence Features and Stability Determinants in Antibody Variable Domains by High-Throughput Experiments
Loop-Sequence Features and Stability Determinants in Antibody Variable Domains by High-Throughput Experiments
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DOI：
10.1016/j.str.2013.10.005
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发表时间：
2014-01-07
期刊：
影响因子：
5.7
通讯作者：
Yang, An-Suei
中科院分区：
文献类型：
--
作者：
Chang, Hung-Ju;Jian, Jhih-Wei;Yang, An-Suei
蛋白质环通常被认为是蛋白质结构和功能的关键决定因素。DNA测序和热稳定性测量的高通量方法的最新进展使得有效地探索局部蛋白质区域的序列-结构-功能关系成为可能。利用这些数据密集型技术，我们研究了模型血管内皮生长因子（VEGF）结合单链抗体可变片段（scFv）可变区域中6个互补决定区（cdr）和10个非cdr环的序列-结构-功能关系，其序列已通过共识序列方法优化。结果表明，只有少数涉及远离抗原结合位点的远程三级相互作用的残基与抗原结合强烈偶联。这表明这些环是蛋白质折叠的被动区域；这些区域的基本序列是由保守的三级相互作用决定的，共识的局部环序列特征对蛋白质的稳定性和功能贡献不大。
Protein loops are frequently considered as critical determinants in protein structure and function. Recent advances in high-throughput methods for DNA sequencing and thermal stability measurement have enabled effective exploration of sequence-structure-function relationships in local protein regions. Using these data-intensive technologies, we investigated the sequence-structure-function relationships of six complementarity-determining regions (CDRs) and ten non-CDR loops in the variable domains of a model vascular endothelial growth factor (VEGF)-binding single-chain antibody variable fragment (scFv) whose sequence had been optimized via a consensus-sequence approach. The results show that only a handful of residues involving long-range tertiary interactions distant from the antigen-binding site are strongly coupled with antigen binding. This implies that the loops are passive regions in protein folding; the essential sequences of these regions are dictated by conserved tertiary interactions and the consensus local loop-sequence features contribute little to protein stability and function.