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Influence of PEGylation on Domain Dynamics of Phosphoglycerate Kinase: PEG Acts Like Entropic Spring for the Protein
Bioconjugate Chemistry ( IF 4.7 ) Pub Date : 2018-05-30 00:00:00 , DOI: 10.1021/acs.bioconjchem.8b00203
Karol Ciepluch 1 , Aurel Radulescu 2 , Ingo Hoffmann 3 , Andreas Raba 1 , Jürgen Allgaier 1 , Dieter Richter 1 , Ralf Biehl 1
Affiliation  

Protein–polymer conjugation is a widely used technique to develop protein therapeutics with improved pharmacokinetic properties as prolonged half-life, higher stability, water solubility, lower immunogenicity, and antigenicity. Combining biochemical methods, small angle scattering (SAXS/SANS), and neutron spin–echo spectroscopy, here we examine the impact of PEGylation (i.e., the covalent conjugation with poly(ethylene glycol) or PEG) on structure and internal domain dynamics of phosphoglycerate kinase (PGK) to elucidate the reason for reduced activity that is connected to PEGylation. PGK is a protein with a hinge motion between the two main domains that is directly related to function. We find that secondary structure and ligand access to the binding sites are not affected. The ligand induced cleft closing is unchanged. We observe an additional internal motion between covalent bonded PEG and the protein compatible with Brownian motion of PGK in a harmonic potential. Entropic interaction with the full PEG chain leads to a force constant of about 8 pN/nm independent of PEG chain length. This additional force preserves protein structure and has negligible effects on the functional domain dynamics of the protein. PEGylation seems to reduce activity just by acting as a local crowder for the ligands. The newly identified interaction mechanism might open possibilities to improve rational design of protein–polymer conjugates.

中文翻译:

聚乙二醇化对磷酸甘油酸激酶结构域动力学的影响:聚乙二醇像蛋白质的熵弹簧一样起作用

蛋白质-聚合物结合是开发具有更长的半衰期,更高的稳定性,水溶性,更低的免疫原性和抗原性的药代动力学特性的蛋白质疗法的广泛使用的技术。结合生化方法,小角度散射(SAXS / SANS)和中子自旋回波光谱学,这里我们研究了聚乙二醇化(即与聚乙二醇或PEG的共价结合)对磷酸甘油酯结构和内部域动力学的影响激酶(PGK)来阐明与PEG化有关的活性降低的原因。PGK是在两个主要结构域之间具有铰链运动的蛋白质,与功能直接相关。我们发现二级结构和配体访问结合位点不受影响。配体引起的裂缝闭合没有改变。我们观察到共价键PEG和与PGK的布朗运动在谐波电位中兼容的蛋白质之间存在额外的内部运动。与完整PEG链的熵相互作用导致独立于PEG链长的力常数约为8 pN / nm。这种额外的作用力保留了蛋白质结构,并且对蛋白质的功能域动力学影响微不足道。聚乙二醇化似乎仅通过充当配体的局部拥挤剂而降低了活性。新近确定的相互作用机制可能为改善蛋白质-聚合物结合物的合理设计开辟可能性。这种额外的作用力保留了蛋白质结构,并且对蛋白质的功能域动力学影响微不足道。聚乙二醇化似乎仅通过充当配体的局部拥挤剂而降低了活性。新近确定的相互作用机制可能为改善蛋白质-聚合物结合物的合理设计开辟可能性。这种额外的作用力保留了蛋白质结构,并且对蛋白质的功能域动力学影响微不足道。聚乙二醇化似乎仅通过充当配体的局部拥挤剂而降低了活性。新近确定的相互作用机制可能为改善蛋白质-聚合物结合物的合理设计开辟可能性。
更新日期:2018-05-30
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