Related Experiment Videos
Volume exclusion effect as a driving force for reverse proteolysis. Implications for polypeptide assemblage in a
Balajee R Somalinga1, Rajendra P Roy
1National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
The Journal of Biological Chemistry
|September 19, 2002
Summary
Macromolecular crowding enhances reverse proteolysis by increasing excluded volume, facilitating polypeptide synthesis and protein complex formation. This effect is significant when protein assembly leads to substantial volume reduction.
Area of Science:
- Biochemistry
- Biophysics
- Molecular Biology
Background:
- Macromolecular crowding influences reactions involving volume changes.
- Reverse proteolysis offers a pathway for protein synthesis and assembly.
Purpose of the Study:
- To investigate the impact of macromolecular crowding on reverse proteolysis.
- To determine the conditions under which crowding facilitates protein formation.
Main Methods:
- Studied subtilisin-catalyzed reformation of triosephosphate isomerase (TIM) from fragments.
- Examined ribonuclease S (RNase S) nick ligation under crowded conditions.
Main Results:
- Crowding facilitated the synthesis of coiled-coil polypeptides.
- Crowding enhanced TIM reformation due to significant volume reduction.
- Crowding did not facilitate RNase S ligation due to minimal volume change.
Conclusions:
- Macromolecular crowding promotes reverse proteolysis when product formation involves significant volume exclusion.
- These findings have implications for in vitro and in vivo protein assembly.