PEGylation of cytochrome c at the level of lysine residues mediated by a microbial transglutaminase
Jian Qin Zhou 1, Ting He 1, Jian Wen Wang 2
Abstract
Objectives: To establish a method for microbial transglutaminase (mTG)-mediated PEGylation of proteins at the level of lysine (Lys) residues.
Results: Carboxybenzyl-glutaminyl-glycinyl-methoxypolyethylene glycol (CBZ-QG-mPEG) was prepared by introducing carboxybenzyl-glutaminyl-glycine (CBZ-QG) to mPEG amine. The analysis by Fourier transform infrared spectroscopy and SDS-PAGE showed that CBZ-QG-mPEG was successfully synthesized and can be recognized by mTG as an acyl donor to modify therapeutic protein, cytochrome c (cyt c). Finally, under an optimized condition (cyt c 0.5 mg/ml, CBZ-QG-mPEG 11.25 mg/ml, mTG 0.5 mg/ml, 37 °C, 2 h), the PEGylation yield reached 76.5 %.
Conclusions: This is the first study regarding the PEGylation of protein at the level of Lys residues catalyzed by mTG. The novel method could be employed to immobilize active proteins and modify therapeutic proteins.
Keywords: Lysine residue; Microbial transglutaminase; PEGylation; Therapeutic protein; Transglutaminase.
Abbreviation: mPEG-NH2
Name: Methoxypoly(ethylene glycol) amine
Abbreviation: mPEG-SPA
Name: Methoxypoly(ethylene glycol) succinimidyl propionate
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