1. Selective Attachment of Polyethylene Glycol to Hemerythrin for Potential Use in Blood Substitutes.
- Author
-
Arkosi, Mariann-Kinga, Mot, Augustin C., Lupan, Iulia, Tegla, Miruna Georgiana Ghinia, and Silaghi-Dumitrescu, Radu
- Subjects
POLYETHYLENE glycol ,BLOOD substitutes ,OXYGEN carriers ,ULTRAVIOLET-visible spectroscopy ,OXIDATION ,SITE-specific mutagenesis ,OXYGEN consumption - Abstract
Due to its ability to reversibly bind O
2 , alongside a relatively low redox reactivity and a limited cytotoxicity, the oxygen-carrying protein hemerythrin has been considered as an alternative to hemoglobin in preparing blood substitutes. In order to increase the hydrodynamic volume and lower antigenicity, two site-directed variants, H82C and K92C, were engineered that contained a single cysteine residue on the surface of each hemerythrin octamer for the specific attachment of polyethylene glycol (PEG). A sulfhydryl-reactive PEGylation reagent with a 51.9 Å spacer arm was used for selective cysteine derivatization. The mutants were characterized by UV-vis spectroscopy, size-exclusion chromatography, oxygen affinity, and autooxidation rate measurements. The H82C variant showed altered oligomeric behavior compared to the wild-type and was unstable in the met form. The PEGylated K92C variant is reasonably stable, displays an oxygen affinity similar to that of the wild-type, and shows an increased rate of autoxidation; the latter disadvantage may be counteracted by further chemical modifications. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF