Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence

FEBS Letters

Tsybovsky, Yaroslav I¹  Martsev, Sergey P¹  Kedrov, Alexey A¹ 

[1] Institute of Bio-Organic Chemistry, National Academy of Sciences of Belarus, Minsk 220141, Belarus

关键词: Immunotoxin;    RNase;    Differential scanning calorimetry;    Protein folding;    Antiferritin;    GMP;    guanosine 3′-monophosphate;    scFv;    single-chain Fv fragment;    VH;    antibody heavy chain variable domain;    VL;    antibody light chain variable domain;   

DOI  :  10.1016/S0014-5793(03)01509-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Although stability is critical for in vivo application of immunotoxins, a thermodynamic description of their folding/stability is still lacking. We applied differential scanning calorimetry (DSC) to RNase-based immunofusion comprising barnase, cytotoxic RNase from Bacillus amyloliquefaciens, fused to the light chain variable domain (VL) of anti-human ferritin antibody F11. By analyzing DSC curves recorded with or without preheating and addition of the barnase-stabilizing ligand guanosine 3′-monophosphate, we (i) assigned two well-resolved thermal transitions to the VL and barnase modules of VL-barnase, (ii) demonstrated independent folding of these two modules, and (iii) showed altered stability of the barnase module, which resulted from the dimeric state of VL-barnase.

【 授权许可】

Unknown