Binding is not enough: flexibility is needed for photocrosslinking of Lck kinase by benzophenone photoligands.
Binding is not enough: flexibility is needed for photocrosslinking of Lck kinase by benzophenone photoligands.
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DOI：
10.1016/j.bmc.2008.08.077
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发表时间：
2008-10-01
影响因子：
3.5
通讯作者：
Aminova O
中科院分区：
文献类型：
--
作者：
Kawamura A;Hindi S;Mihai DM;James L;Aminova O
二苯甲酮类光生团被广泛用于光亲和标记研究。然而，用二苯甲酮进行光标记很难说是一项常规实验。即使当光电探测器结合到它的靶上时，光交联也不一定发生。这是因为除了靶结合外，二苯甲酮的光标记还受到许多因素的影响，例如光配体的构象灵活性。尽管人们普遍认识到这种并发症，但还没有系统的研究来评估影响光标记效率的单个因素的相对重要性。为了深入了解这一问题，我们进行了二苯甲酮类光配体与LCK激酶的构效关系(SAR)研究，比较了不同靶向结合亲和力和构象柔性的光配体。这项研究发现，结合亲和力与光标记效率无关，这一结合亲和力由激酶抑制效力表示。相反，构象灵活性被发现是我们的光配体有效光标记的决定因素。讨论了当前发现的意义，特别是关于二苯甲酮光配体的选择和优化。
Benzophenone photophores are employed widely for photoaffinity-labeling studies. Photolabeling with benzophenone, however, is hardly a routine experiment. Even when a photoprobe binds to its target, photocrosslinking does not necessarily occur. This is because photolabeling by benzophenone is affected by many factors other than target-binding, such as conformational flexibility of photoligand. Despite the widespread recognition of such complications, there has been no systematic study to assess the relative importance of individual factors that can affect photolabeling efficiency. In order to gain an insight into this problem, we conducted a structure-activity relationship (SAR) study of benzophenone photoligands for Lck kinase, in which photoligands with varying target binding-affinity and conformational flexibility were compared. The study found that binding-affinity, as indicated by kinase inhibitory potency, did not correlate with photolabeling efficiency. Instead, conformational flexibility was found to be the determining factor for efficient photolabeling by our photoligands. Implication of the current findings, in particular, with regard to selection and optimization of benzophenone photoligands, is discussed.
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影响因子：
64.8
作者：
ANDERSON, SJ;LEVIN, SD;PERLMUTTER, RM
通讯作者：
PERLMUTTER, RM
影响因子：
3.3
作者：
Prestwich, GD;Dorman, G;Chaudhary, A
通讯作者：
Chaudhary, A
影响因子：
15.9
作者：
Goldman, FD;Ballas, ZK;Taylor, N
通讯作者：
Taylor, N
DOI：
10.1073/pnas.0608659104
发表时间：
2007-01-23
影响因子：
11.1
作者：
Salisbury, Cleo M.;Cravatt, Benjamin F.
通讯作者：
Cravatt, Benjamin F.
影响因子：
15
作者：
Chan, EWS;Chattopadhaya, S;Yao, SQ
通讯作者：
Yao, SQ