CDMS Analysis of Intact 19S, 20S, 26S, and 30S Proteasomes: Evidence for Higher-Order 20S Assemblies at a Low pH†.
CDMS Analysis of Intact 19S, 20S, 26S, and 30S Proteasomes: Evidence for Higher-Order 20S Assemblies at a Low pH†.
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完整 19S、20S、26S 和 30S 蛋白酶体的 CDMS 分析：低 pH 下高阶 20S 组装的证据。
DOI：
10.1021/acs.analchem.3c00472
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发表时间：
2023
影响因子：
7.4
通讯作者：
Clemmer,DavidE
中科院分区：
文献类型：
--
作者：
Anthony,AdamJ;Gautam,AmitKS;Miller,LohraM;Ma,Yiran;Hardwick,AnyaG;Sharma,Anu;Ghatak,Subhadip;Matouschek,Andreas;Jarrold,MartinF;Clemmer,DavidE
电荷检测质谱（CDMS）是研究蛋白酶体的一种手段。为此，测定了酿酒酵母20S、19S、26S和30S蛋白酶体的质量：m(20S) = 738.8±2.9 kDa,m(19S) = 926.2±4.8 kDa,m(26S) = 1637.0±7.6 kDa,m(30S) = 2534.2±10.8 kDa。在某些条件下，观察到较大的（20S）x（其中x= 1至~ 13）组装；19S调节颗粒也发生寡聚，但在较小程度上形成（19S）x复合物（其中x= 1至4，有利于x= 3三聚体）。（20S）低聚物在体外更受青睐，因为溶液的pH值降低（在20 mM醋酸铵溶液中从7.0降至5.4），这可能与体内在碳饥饿下观察到的蛋白酶体储存颗粒有关。从m(20S)x（x= 1到~ 13）个物种的测量中，似乎每个多聚体保留了20S复合物亚基的所有28种蛋白质。考虑了几种可能解释（20S）xassemblies形成的结构类型。我们强调每种结构类型[假设平面，筏状几何形状（其中单个蛋白酶体通过并排相互作用相关联）；细长的棒状几何形状（其中亚单元端到端绑定）；大致为球形的几何形状（通过非特异性亚基相互作用产生的聚集）是高度推测性的，但考虑起来仍然很有趣，并提供了简短的讨论。讨论了CDMS在表征蛋白酶体和相关低聚物方面的应用。
Charge detection mass spectrometry (CDMS) was examined as a means of studying proteasomes. To this end, the following masses of the 20S, 19S, 26S, and 30S proteasomes fromSaccharomyces cerevisiae(budding yeast) were measured:m(20S) = 738.8 ± 2.9 kDa,m(19S) = 926.2 ± 4.8 kDa,m(26S) = 1,637.0 ± 7.6 kDa, andm(30S) = 2,534.2 ± 10.8 kDa. Under some conditions, larger (20S)x(wherex= 1 to ∼13) assemblies are observed; the 19S regulatory particle also oligomerizes, but to a lesser extent, forming (19S)xcomplexes (wherex= 1 to 4, favoring thex= 3 trimer). The (20S)xoligomers are favored in vitro, as the pH of the solution is lowered (from 7.0 to 5.4, in a 20 mM ammonium acetate solution) and may be related to in vivo proteasome storage granules that are observed under carbon starvation. From measurements ofm(20S)x(x= 1 to ∼13) species, it appears that each multimer retains all 28 proteins of the 20S complex subunit. Several types of structures that might explain the formation of (20S)xassemblies are considered. We stress that each structural type [hypothetical planar, raft-like geometries (where individual proteasomes associate through side-by-side interactions); elongated, rodlike geometries (where subunits are bound end-to-end); and geometries that are roughly spherical (arising from aggregation through nonspecific subunit interactions)] is highly speculative but still interesting to consider, and a short discussion is provided. The utility of CDMS for characterizing proteasomes and related oligomers is discussed.
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