Backbone assignment of a 28.5 kDa class A extended spectrum β-lactamase by high-field, carbon-detected solid-state NMR.

IF 0.6 4区 生物学 Q4 BIOPHYSICS
Christopher G Williams, Songlin Wang, Alexander F Thome, Owen A Warmuth, Varun Sakhrani, Leighton Coates, Chad M Rienstra, Leonard J Mueller
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引用次数: 0

Abstract

13C and 15N backbone chemical shift assignments are reported for the 28.5 kDa protein Toho-1 β-lactamase, a Class A extended spectrum β-lactamase. A very high level of assignment completeness (97% of the backbone) is enabled by the combined sensitivity and resolution gains of ultrahigh-field NMR spectroscopy (1.1 GHz), improved probe technology, and optimized pulse sequences. The assigned chemical shifts agree well with our previous solution-state NMR assignments, indicating that the secondary structure is conserved in the solid state. These assignments provide a foundation for future investigations of side-chain chemical shifts and catalytic mechanism.

一个28.5 kDa的a类扩展光谱β-内酰胺酶的高场碳检测固体核磁共振骨架分配。
报道了28.5 kDa蛋白Toho-1 β-内酰胺酶(a类扩展谱β-内酰胺酶)的13C和15N主链化学位移定位。通过超高场核磁共振波谱(1.1 GHz)的灵敏度和分辨率增益、改进的探针技术和优化的脉冲序列,可以实现非常高水平的分配完整性(97%的骨干)。分配的化学位移与我们之前的溶液态NMR分配非常一致,表明二级结构在固态中是守恒的。这些作业为今后研究侧链化学位移和催化机理提供了基础。
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来源期刊
Biomolecular NMR Assignments
Biomolecular NMR Assignments 生物-光谱学
CiteScore
1.70
自引率
11.10%
发文量
59
审稿时长
6-12 weeks
期刊介绍: Biomolecular NMR Assignments provides a forum for publishing sequence-specific resonance assignments for proteins and nucleic acids as Assignment Notes. Chemical shifts for NMR-active nuclei in macromolecules contain detailed information on molecular conformation and properties. Publication of resonance assignments in Biomolecular NMR Assignments ensures that these data are deposited into a public database at BioMagResBank (BMRB; http://www.bmrb.wisc.edu/), where they are available to other researchers. Coverage includes proteins and nucleic acids; Assignment Notes are processed for rapid online publication and are published in biannual online editions in June and December.
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