Quantitative analysis of dissociation of LDH by high pressure native PAGE

ABSTRACT

High pressure native polyacrylamide gel electrophoresis has been designed to visualize the dissociation/association process of protein complexes. This paper reports this methodology in more quantitative way by inspecting pressure dissociation of pig heart lactate dehydrogenase, a tetrameric protein, which was extensively investigated in spectroscopic methods. We observed the change of electrophoresis pattern with pressure up to 150 MPa. By optimizing the buffer system and careful image analysis of the stained gels, we quantified all the dissociates in the process of pressurization. We discussed the characteristics of our methodology by comparing the result with the previously reported.

KEYWORDS:

• High pressure native PAGE
• Ferguson plot analysis
• lactate dehydrogenase
• oligomer dissociation

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by JSPS KAKENHI [grant number JP16K05814].