Electromanipulation of proteins using nanosecond pulsed electric fields

Abstract

The present disclosure describes methods for intracellular electromanipulation of proteins using nanosecond pulsed electric fields (nsPEFs). The nsPEFs have effects on proteins in addition to permeabilizing cellular membranes. The nsPEFs induce a Ca2+-dependent dissipation of the mitochondria membrane potential (ΔΨm), which is enhanced when high frequency components are present in fast rise-fall waveforms. Ca2+ is shown to have little or no effect on propidium iodide uptake as a measure of plasma membrane poration and consequently intracellular membranes. Since Ca2+-regulated events are mediated by proteins, actions of nsPEFs on proteins that regulate and/or affect the mitochondria membrane potential are possible. Given that nsPEF-induced dissipation of ΔΨm was more effective when high frequency components were present in fast rise time waveforms, the effects on proteins are due to these high frequency components. These results present direct evidence that nsPEFs affect proteins and their functions by affecting their structure.

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