Intense Pulsed Electric Fields Denature Urease Protein.

Area of Science:

• Biophysics
• Biochemistry
• Protein Science

Background:

• Nanosecond pulsed electric fields (nsPEFs) are increasingly studied for their biological effects.
• Understanding protein response to nsPEFs is crucial for applications in biotechnology and medicine.
• Previous research has explored nsPEF interactions with biological molecules, but protein-specific structural and functional impacts require further elucidation.

Purpose of the Study:

• To investigate the effects of high-intensity nsPEFs on the structure and enzyme activity of specific proteins.
• To determine if nsPEFs induce structural changes in lysozyme, albumin, and urease.
• To assess the impact of nsPEFs on the enzymatic function of urease.

Main Methods:

• Solutions of lysozyme, albumin, and urease were subjected to intense nsPEFs (up to 300 kV/cm, 5-ns duration, 500 pulses at 3 Hz).
• Structural analysis focused on covalent bonds (peptide, disulfide) for lysozyme and albumin.
• Tertiary and quaternary structures, along with enzyme activity, were analyzed for urease.

Main Results:

• High-intensity nsPEFs (≥250 kV/cm) caused deformation in the quaternary and tertiary structures of urease.
• Urease enzyme activity decreased significantly at field strengths of 250 kV/cm and higher.
• No significant structural changes were observed in lysozyme or albumin, even at the maximum field strength of 300 kV/cm.

Conclusions:

• Intense nsPEFs can physically alter the conformation and function of certain proteins, exemplified by urease.
• Protein susceptibility to nsPEF-induced structural changes varies, with urease showing sensitivity while lysozyme and albumin did not.
• These findings contribute to understanding the physical interactions between intense electric fields and protein molecules, relevant to cellular membrane electroporation phenomena.