Gragerov A, Zeng L, Zhao X, Burkholder W, Gottesman M E
Institute of Cancer Research, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
J Mol Biol. 1994 Jan 21;235(3):848-54. doi: 10.1006/jmbi.1994.1043.
The sequence specificity of DnaK substrate binding has been studied using a peptide display library. Based on the amino acid patterns that appeared in this selection, short peptides were synthesized for direct measurements of DnaK affinity. The results show that peptides enriched in internal hydrophobic residues are preferential DnaK substrates, and negatively charged peptides have poor affinity. The isolated C-terminal domain of DnaK binds peptides. Peptide dissociation studies indicate that bound peptides are released from the C-terminal fragment and from DnaK at identical rates. ATP stimulates peptide dissociation from DnaK but not from the C-terminal fragment.
已使用肽展示文库研究了DnaK底物结合的序列特异性。基于该筛选中出现的氨基酸模式，合成了短肽以直接测量DnaK亲和力。结果表明，富含内部疏水残基的肽是优先的DnaK底物，而带负电荷的肽亲和力较差。分离出的DnaK C末端结构域可结合肽。肽解离研究表明，结合的肽以相同速率从C末端片段和DnaK中释放。ATP刺激肽从DnaK解离，但不刺激从C末端片段解离。
