A shift in the salt bridge interaction of residues D620 and E621 mediates the constitutive activation of Jak2-H538Q/K539L

被引:4
作者
Gnanasambandan, Kavitha [1 ]
Magis, Andrew T. [2 ]
Sayeski, Peter P. [1 ]
机构
[1] Univ Florida, Coll Med, Dept Physiol & Funct Genom, Gainesville, FL 32610 USA
[2] Univ Illinois, Ctr Biophys & Computat Biol, Urbana, IL 61801 USA
基金
美国国家卫生研究院;
关键词
Jak2; Exon; 12; mutation; MPNs; Salt bridge; Simulations; Mechanism; TYROSINE KINASE JAK2; MOLECULAR-DYNAMICS; POLYCYTHEMIA-VERA; PROTEIN-KINASE; MUTATION; INHIBITORS; MECHANISM; FUSION;
D O I
10.1007/s11010-012-1326-7
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Jak2 mutations in the exon 14 and exon 12 regions that cause constitutive activation have been associated with myeloproliferative neoplasms. We have previously shown that a pi stacking interaction between F617 and F595 is important for the constitutive activation of Jak2-V617F (Gnanasambandan et al., Biochemistry 49:9972-9984, 2010). Here, using a combination of molecular dynamics (MD) simulations and in vitro mutagenesis, we studied the molecular mechanism for the constitutive activation of the Jak2 exon 12 mutation, H538Q/K539L. The activation levels of Jak2-H538Q/K539L were found to be similar to that of Jak2-V617F, and Jak2-H538Q/K539L/V617F. Data from MD simulations indicated a shift in the salt bridge interactions of D620 and E621 with K539 in Jak2-WT to R541 in Jak2-H538Q/K539L. When compared to Jak2-WT, K539A mutation resulted in increased activation, while K539D or K539E mutations diminished Jak2 activation by 50 %. In the context of Jak2-H538Q/K539L, R541A mutation reduced its activation by 50 %, while R541D and R541E mutations returned its activation levels to that of Jak2-WT. Collectively, these results indicate that a shift in the salt bridge interaction of D620 and E621 with K539 in Jak2-WT to R541 in Jak2-H538Q/K539L is critical for constitutive activation of this Jak2 exon 12 mutant.
引用
收藏
页码:125 / 140
页数:16
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