allosteric interactions between the myristate- and atp-site of the abl kinase变构之间的交互的豆蔻酸盐和atp-site abl激酶.pdfVIP

Allosteric Interactions between the Myristate- and ATP- Site of the Abl Kinase 1¤ 2 2 1 Roxana E. Iacob , Jianming Zhang , Nathanael S. Gray , John R. Engen * 1 Department of Chemistry Chemical Biology, and The Barnett Institute for Chemical and Biological Analysis, Northeastern University, Boston, Massachusetts, United States of America, 2 Biological Chemistry Molecular Pharmacology, Harvard Medical School and Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, United States of America Abstract Abl kinase inhibitors targeting the ATP binding pocket are currently employed as potent anti-leukemogenic agents but drug resistance has become a significant clinical limitation. Recently, a compound that binds to the myristate pocket of Abl (GNF-5) was shown to act cooperatively with nilotinib, an ATP-competitive inhibitor to target the recalcitrant ‘‘T315I’’ gatekeeper mutant of Bcr-Abl. To uncover an explanation for how drug binding at a distance from the kinase active site could lead to inhibition and how inhibitors could combine their effects, hydrogen exchange mass spectrometry (HX MS) was employed to monitor conformational effects in the presence of both dasatinib, a clinically approved ATP-site inhibitor, and GNF-5. While dasatinib binding to wild type Abl clearly influenced Abl conformation, no binding was detected between dasatinib and T315I. GNF-5, however, elicited the same conformational changes in both wild type and T315I, including ˚ changes to dynamics within the ATP site l