NMR structure calculation 1
NMR structure calculation 1
NMR结构解析流程 生物信息学和生物化学分析 样品制备（蛋白质表达纯化，标记） 核磁共振初步鉴定 ↓ 核磁共振数据收集 ] 化学位移指认二 二级结构分析 ↓ NOE指认 结构计算 2
NMR结构解析流程 生物信息学和生物化学分析 样品制备（蛋白质表达纯化，标记） 核磁共振数据收集 化学位移指认 NOE指认 结构计算 核磁共振初步鉴定 二级结构分析 2
Solving structures by NMR Sample Preparation Structural restraints .Cloning,expression,purification .NOE,H-bonds .Isotope labelling [5N,13C/15N,[2H/3C/15N ·J-couplings .Residual dipolar couplings,T1/T2 .Chemical shifts Resonance Assignments ·Backbone ·Side chains Structure Calculation .Distance geometry .Restrained molecular dynamics .Simulated annealing Secondary Structure Chemical shift Ensemble of3D structures 3
Solving structures by NMR Sample Preparation •Cloning, expression, purification •Isotope labelling [ 15N], [13C/15N],[ 2H/13C/15N] Resonance Assignments • Backbone • Side chains Secondary Structure Chemical shift Structural restraints •NOE, H-bonds •J-couplings •Residual dipolar couplings, T1/T2 •Chemical shifts Structure Calculation •Distance geometry •Restrained molecular dynamics •Simulated annealing Ensemble of 3D structures 3
Overview Structure representation Types of NMR data conversion into restraints Structure calculation methods Structure validation 4
Overview • Structure representation • Types of NMR data conversion into restraints • Structure calculation methods • Structure validation 4
a structure 蛋白质结构层次 氨基酸通过肽键形成的生物高分子 一级结构、二级结构、三级结构、四级结构 econdary structure i-1 i+1 R:侧链 R R RO N℃℃N=CN=C0 B pleated sheet 主链 ..出. alpha helix •肽键具有双键性质而不能任意旋转 主链可旋转的二面角φ，w tertiary structure 15 quaternary structure N-C C-N
蛋白质结构层次 • 氨基酸通过肽键形成的生物高分子 • 一级结构、二级结构、三级结构、四级结构 •肽键具有双键性质而不能任意旋转 •主链可旋转的二面角， 5