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Independent folding and conformational changes of the barnase module in the VL‐barnase immunofusion: calorimetric evidence
Author(s) -
Tsybovsky Yaroslav I,
Kedrov Alexey A,
Martsev Sergey P
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)01509-6
Subject(s) - barnase , differential scanning calorimetry , bacillus amyloliquefaciens , rnase p , chemistry , crystallography , folding (dsp implementation) , biophysics , biochemistry , thermodynamics , ribonuclease , biology , physics , rna , fermentation , electrical engineering , gene , engineering
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.