Cryptophane xenon-129 nuclear magnetic resonance biosensors targeting human carbonic anhydrase.
Cryptophane xenon-129 nuclear magnetic resonance biosensors targeting human carbonic anhydrase.
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DOI：
10.1021/ja806092w
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发表时间：
2009-01-21
影响因子：
15
通讯作者：
Dmochowski, Ivan J.
中科院分区：
文献类型：
--
作者：
Chambers, Jennifer M.;Hill, P. Aru;Aaron, Julie A.;Han, Zhaohui;Christianson, David W.;Kuzma, Nicholas N.;Dmochowski, Ivan J.
129 NMR生物传感器是有前途的试剂，用于早期检测疾病，因为生物分子相互作用可以干扰129 NMR化学位移远远超出临床MRI的典型场不均匀性。我们介绍了人类碳酸酐酶（CA）作为一个单一的结合位点的酶研究氙生物传感器蛋白质的相互作用。一个氙结合的cryptophane取代与连接器的不同长度的对苯磺酰胺，以产生非非对映异构体的生物传感器与一个单一的129 NMR共振。X射线晶体学证实了在CAII活性位点中含有单个氙原子的8键连接的生物传感器的结合。通过等温滴定量热法（ITC）测定同工酶CA I和II的生物传感器解离常数（Kd = 20-110 nM）。生物传感器-CA复合物产生了窄线宽的“结合”超极化129 NMR共振，其向低场移动了3.0至7.5 ppm，这意味着比以前看到的大得多的位移。此外，同工酶特异性的化学位移明显区分CA I和II，尽管他们相似的活性位点架构。因此，氙生物传感器可以提供一个强大的战略，用于诊断人类疾病的特点是特定的CA同工酶和其他蛋白质生物标志物的上调。
129Xe NMR biosensors are promising agents for the early detection of diseases, given that biomolecular interactions can perturb 129Xe chemical shifts well beyond the typical field inhomogeneity of clinical MRI. We introduce human carbonic anhydrase (CA) as a single-binding-site enzyme for studying xenon biosensor-protein interactions. A xenon-binding cryptophane was substituted with linkers of varying lengths to p-benzenesulfonamide to yield non-diastereomeric biosensors with a single 129Xe NMR resonance. X-ray crystallography confirmed binding of the 8-bond-linked biosensor containing a single xenon atom in the CAII active site. Biosensor dissociation constants (Kd = 20–110 nM) were determined by isothermal titration calorimetry (ITC) for isozymes CA I and II. The biosensor-CA complexes yielded “bound” hyperpolarized 129Xe NMR resonances of narrow linewidth that were shifted by 3.0 to 7.5 ppm downfield, signifying much larger shifts than seen previously. Moreover, isozyme-specific chemical shifts clearly differentiated CA I and II, despite their similar active-site architectures. Thus, xenon biosensors may provide a powerful strategy for diagnosing human diseases characterized by the upregulation of specific CA isozymes and other protein biomarkers.
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影响因子：
3.3
作者：
Foster-Gareau, P;Heyn, C;Rutt, BK
通讯作者：
Rutt, BK
影响因子：
3.3
作者：
Mugler, JP;Driehuys, B;Wagshul, ME
通讯作者：
Wagshul, ME
影响因子：
3.2
作者：
Lowery, TJ;Garcia, S;Wemmer, DE
通讯作者：
Wemmer, DE
影响因子：
5
作者：
Broome, Dale R.;Girguis, Mark S.;Kirk, Gerald A.
通讯作者：
Kirk, Gerald A.
影响因子：
64.8
作者：
ALBERT, MS;CATES, GD;WISHNIA, A
通讯作者：
WISHNIA, A