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| Modifiation of Cytochrome c at the Level of Lysine Residues Mediated by Microbial Transglutaminase |
| ZHANG Chen1, CHEN Shao-hua2, WU Wen-qian2, ZHOU Jian-qin1 |
1. College of Pharmaceutical Sciences, Soochow University, Suzhou 215123, China;
2. Experimental Center of Medical College, Soochow University, Suzhou 215123, China |
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Abstract Experiments were carried out to investigate the possibilities of site specific PEGylation of therapeutic proteins (cytochrome c, Cyt c) catalyzed by mTG.Then reaction conditions were optimized and the properties of the PEGylated Cyt c were explored. CBZ-QG-mPEG was successfully synthesized by introducing CBZ-QG into methoxypolyethylene glycol amine (mPEG-NH2) and could act as the acyl donor of mTG.The possibilities of mPEG-NH2 acting as an amino donor or CBZ-QG-mPEGacting as an acyl donor to modify Cyt c catalyzed by mTG were investigated. CBZ-QG-mPEG was coupled to the specific Lys residue of Cyt c catalyzed by mTG.The optimized PEGylationconditions were as follows:37℃, pH 8.0, mTG 1.0mg/ml, reaction time 2h. The PEGylated Cyt c exhibited better thermal stability and pH stability than the native Cyt c. Only one specific Lys residue of Cyt c was PEGylated catalyzed by mTG, while several Lys residues of Cyt c were conjugated with mPEG-SPA by using the chemical method, which leading to the heterogeneity of the derivatives. The prominent advantage of mTG-mediated catalysis is its highsite specificity and thus homogeneity.Here, the mTG-mediated PEGylation of proteins at the level of lysine (Lys) residues was developed, which overcome the random modification of Lys residues by the chemical method andwas expected to be generally applied to protein modification.
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Received: 14 February 2017
Published: 25 September 2017
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