An in vitro peptide folding model suggests the presence of the molten globule state during nascent peptide folding.
Although molten globules have been widely accepted as a general intermediate in protein folding, there is no clear evidence to show their presence during nascent peptide folding. This paper concentrates on whether the molten globule state occurs, and if it does, when does it form during nascent peptide folding, by comparing the changes in conformation during peptide chain extension of staphylococcal nuclease R. The results show that a large N-terminal fragment of staphylococcal nuclease, SNR121, which already contains more than 80% amino acid sequence of the nuclease, is found to fulfill all the criteria for the molten globule state, suggesting that the molten globule should occur at a later stage of peptide elongation. At this stage the hydrophobic collapse of the polypeptide chain occurs driven by the hydrophobic force, which leads to the formation of a solvent-accessible non-polar core, characterized by the high ANS-binding fluorescence. The nascent peptide folding of the nuclease is a hierarchical process that at the very least includes the following steps: secondary structure accumulation, pre-molten globule state, molten globule state, post-molten globule state and finally the native state. Constant conformation adjustment is necessary for correct folding and active expression of the protein.
Zhou
B
B
National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing 100101, People's Republic of China.
Tian
K
K
Jing
G
G
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry / Q000378:metabolism; D002942:Circular Dichroism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D010446:Peptide Fragments; C027132:1-anilino-8-naphthalenesulfonate; C104574:staphylococcal nuclease R; D008836:Micrococcal Nuclease