PUBLICATION
Improving the cell-membrane-penetrating activity of globins by introducing positive charges on protein surface: A case study of sperm whale myoglobin
- Authors
- Gao, S.Q., Yuan, H., Yang, X.Z., Xiang, H.F., Tan, X., Wen, G.B., Lin, Y.W.
- ID
- ZDB-PUB-220213-12
- Date
- 2022
- Source
- Biochemical and Biophysical Research Communications 598: 26-31 (Journal)
- Registered Authors
- Keywords
- Cell membrane, Globins, Heme proteins, Protein transduction, X-ray crystallography
- MeSH Terms
-
- MCF-7 Cells
- Sperm Whale
- Spectrophotometry, Ultraviolet
- Fluorescein-5-isothiocyanate/chemistry
- Animals
- Cell Membrane/drug effects
- Cell Membrane/metabolism*
- Lysine/chemistry
- Crystallography, X-Ray
- Myoglobin/chemistry*
- Myoglobin/genetics
- Myoglobin/metabolism*
- Myoglobin/pharmacokinetics
- Mutation
- Cell Survival/drug effects
- Circular Dichroism
- Humans
- PubMed
- 35151200 Full text @ Biochem. Biophys. Res. Commun.
Citation
Gao, S.Q., Yuan, H., Yang, X.Z., Xiang, H.F., Tan, X., Wen, G.B., Lin, Y.W. (2022) Improving the cell-membrane-penetrating activity of globins by introducing positive charges on protein surface: A case study of sperm whale myoglobin. Biochemical and Biophysical Research Communications. 598:26-31.
Abstract
Globins are heme proteins such as hemoglobin (Hb), myoglobin (Mb) and neuroglobin (Ngb), playing important roles in biological system. In addition to normal functions, zebrafish Ngb was able to penetrate cell membranes, whereas less was known for other globin members. In this study, to improve the cell-membrane-penetrating activity of globins, we used sperm whale Mb as a model protein and constructed a quadruple mutant of G5K/Q8K/A19K/V21K Mb (termed 4K Mb), by introduction of four positive charges on the protein surface, which was designed according to the amino acid alignment with that of zebrafish Ngb. Spectroscopic and crystallographic studies showed that the four positively charged Lys residues did not affect the protein structure. Cell-membrane-penetrating essay further showed that 4K Mb exhibited enhanced activity compared to that of native Mb. This study provides valuable information for the effect of distribution of charged residues on the protein structure and the cell-membrane-penetrating activity of globins. Therefore, it will guide the design of protein-based biomaterials for biological applications.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping