Publication Abstracts

Bulos et al. 2021

Bulos, J.A., R. Guo, Z. Wang, M.A. DeLessio, J.G. Saven, and I.J. Dmochowski, 2021: Design of a superpositively charged enzyme: Human carbonic anhydrase II variant with ferritin encapsulation and immobilization. Biochemistry, 60, 3596-3609, doi:10.1021/acs.biochem.1c00515.

Supercharged proteins exhibit high solubility and other desirable properties, but no engineered superpositively charged enzymes have previously been made. Superpositively charged variants of proteins such as green fluorescent protein have been efficiently encapsulated within Archaeoglobus fulgidus thermophilic ferritin (AfFtn). Encapsulation by supramolecular ferritin can yield systems with a variety of sequestered cargo. To advance applications in enzymology and green chemistry, we sought a general method for supercharging an enzyme that retains activity and is compatible with AfFtn encapsulation. The zinc metalloenzyme human carbonic anhydrase II (hCAII) is an attractive encapsulation target based on its hydrolytic activity and physiologic conversion of carbon dioxide to bicarbonate. A computationally designed variant of hCAII contains positively charged residues substituted at 19 sites on the protein's surface, resulting in a shift of the putative net charge from -1 to +21. This designed hCAII(+21) exhibits encapsulation within AfFtn without the need for fusion partners or additional reagents. The hCAII(+21) variant retains esterase activity comparable to the wild type and spontaneously templates the assembly of AfFtn 24mers around itself. The AfFtn-hCAII(+21) host-guest complex exhibits both greater activity and thermal stability when compared to hCAII(+21). Upon immobilization on a solid support, AfFtn-hCAII(+21) retains enzymatic activity and exhibits an enhancement of activity at elevated temperatures. %AB Supercharged proteins exhibit high solubility and other desirable properties, but no engineered superpositively charged enzymes have previously been made. Superpositively charged variants of proteins such as green fluorescent protein have been efficiently encapsulated within Archaeoglobus fulgidus thermophilic ferritin (AfFtn). Encapsulation by supramolecular ferritin can yield systems with a variety of sequestered cargo. To advance applications in enzymology and green chemistry, we sought a general method for supercharging an enzyme that retains activity and is compatible with AfFtn encapsulation. The zinc metalloenzyme human carbonic anhydrase II (hCAII) is an attractive encapsulation target based on its hydrolytic activity and physiologic conversion of carbon dioxide to bicarbonate. A computationally designed variant of hCAII contains positively charged residues substituted at 19 sites on the protein's surface, resulting in a shift of the putative net charge from -1 to +21. This designed hCAII(+21) exhibits encapsulation within AfFtn without the need for fusion partners or additional reagents. The hCAII(+21) variant retains esterase activity comparable to the wild type and spontaneously templates the assembly of AfFtn 24mers around itself. The AfFtn-hCAII(+21) host-guest complex exhibits both greater activity and thermal stability when compared to hCAII(+21). Upon immobilization on a solid support, AfFtn-hCAII(+21) retains enzymatic activity and exhibits an enhancement of activity at elevated temperatures. %M3 doi: 10.1021/acs.biochem.1c00515

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BibTeX Citation

@article{bu01200v,
  author={Bulos, J. A. and Guo, R. and Wang, Z. and DeLessio, M. A. and Saven, J. G. and Dmochowski, I. J.},
  title={Design of a superpositively charged enzyme: Human carbonic anhydrase II variant with ferritin encapsulation and immobilization},
  year={2021},
  journal={Biochemistry},
  volume={60},
  pages={3596--3609},
  doi={10.1021/acs.biochem.1c00515},
  isbn={0006-2960}
}

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RIS Citation

TY  - JOUR
ID  - bu01200v
AU  - Bulos, J. A.
AU  - Guo, R.
AU  - Wang, Z.
AU  - DeLessio, M. A.
AU  - Saven, J. G.
AU  - Dmochowski, I. J.
PY  - 2021
TI  - Design of a superpositively charged enzyme: Human carbonic anhydrase II variant with ferritin encapsulation and immobilization
JA  - Biochemistry
VL  - 60
SP  - 3596
EP  - 3609
DO  - 10.1021/acs.biochem.1c00515
SN  - 0006-2960
ER  -

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