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首页> 外文期刊>Journal of mass spectrometry: JMS >Detection and structural features of the beta B2-B3-crystallin heterodimer by radical probe mass spectrometry (RP-MS)
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Detection and structural features of the beta B2-B3-crystallin heterodimer by radical probe mass spectrometry (RP-MS)

机译：自由基探针质谱法（RP-MS）检测βB2-B3-晶体蛋白异二聚体的结构特征

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摘要

The predilection of the beta-crystallin B2 subunit to interact with the beta B3 subunit rather than self associate is evident by the detection of the beta B2-B3-crystallin heterodimer by native gel electrophoresis and electrospray ionisation time-of-flight (ESI-TOF) mass spectrometry under non denaturing conditions. The complex has been detected for the first time and its molecular mass is measured to be 47 450 +/- 1 Da. Radical probe mass spectrometry (RP-MS) was subsequently applied to investigate the nature of the heterodimer through the limited oxidation of the subunits in the complex. Two peptide segments of the beta B2 subunit and six of the beta B3 subunit were found to oxidise, with far greater oxidation observed within the beta B3 versus the beta B2 subunit. This, and the observation that the oxidation data of beta B2 subunit is inconsistent with the structure of the beta B2 monomer, demonstrates that the protection of beta B2 is conferred by its association with beta B3 subunit within the heterodimer where only the residues of, and towards, its N-terminal domain remain exposed to solvent. The results suggest that the beta B2 subunit adopts a more compacted form than in its monomeric form in order for much of its structure to be enveloped by the beta B3 subunit within the heterodimer.
机译：通过天然凝胶电泳和电喷雾电离飞行时间（ESI-TOF）检测到βB2-B3-晶状蛋白异源二聚体，证明了β-晶状蛋白B2亚基与βB3亚基相互作用而不是自身缔合的偏好。 ）在非变性条件下进行质谱分析。首次检测到该配合物，其分子量为47 450 +/- 1 Da。随后应用了自由基探针质谱（RP-MS），通过对复合物中亚基的有限氧化来研究异二聚体的性质。发现βB2亚基的两个肽段和βB3亚基的六个肽段被氧化，与βB2亚基相比，βB3内的氧化作用更大。这以及βB2亚基的氧化数据与βB2单体的结构不一致的观察结果表明，βB2的保护是由于其与异二聚体中的βB3亚基缔合而赋予的，该异二聚体中仅存在残基，且朝向，其N-末端结构域保持暴露于溶剂。结果表明，βB2亚单位采用比其单体形式更紧密的形式，以便其大部分结构被异二聚体中的βB3亚单位所包围。

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