Conformational stability of HPr: The histidine‐containing phosphocarrier protein from Bacillus subtilis
Conformational stability of HPr: The histidine‐containing phosphocarrier protein from Bacillus subtilis
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HPr 的构象稳定性：来自枯草芽孢杆菌的含组氨酸磷酸载体蛋白
DOI：
10.1002/pro.5560040106
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发表时间：
1995
期刊：
影响因子：
8
通讯作者：
M. Scholtz
中科院分区：
文献类型：
--
作者：
J.;M. Scholtz
采用热解折叠和溶剂变性实验相结合的方法，测定了枯草芽孢杆菌含组氨酸磷酸载体蛋白（HPr）的构象稳定性。在固定温度下用光谱法监测尿素诱导的HPr变性，并在固定浓度的尿素存在下进行热解折叠。这些数据以几种不同的方式进行分析，以提供描述HPr折叠热力学的基本参数（ΔHg，Tg，ΔSg和ΔCp）的测量。使用Pace和Laurents的方法（Pace CN，Laurents DV，1989，Biochemistry 28：2520-2525）估算ΔCP，这是热和尿素诱导的解折叠数据的整体分析。用于分析数据的每种方法都给出了类似的ΔCp值（1，170 Δ 50 cal mol−1 K−1）。尽管HPr的熔融温度很高（Tg = 73.5 °C），但蛋白质的最大稳定性（26 °C）相当适中（AGs = 4.2 kcal mol−1）。在中等浓度的尿素存在下，HPr表现出冷变性，因此使用Chen和Schellman的方法可以获得HPr的完整稳定性曲线，包括ΔCp的测量（Chen B，Schellman JA，1989，Biochemistry 28：685-691）。比较了不同的溶剂变性曲线分析方法，并讨论了尿素对这种小球蛋白热稳定性的影响。所提出的方法将在许多小蛋白质的稳定性曲线的表征中具有通用性。
The conformational stability of the histidine‐containing phosphocarrier protein (HPr) from Bacillus subtilis has been determined using a combination of thermal unfolding and solvent denaturation experiments. The urea‐induced denaturation of HPr was monitored spectroscopically at fixed temperatures and thermal unfolding was performed in the presence of fixed concentrations of urea. These data were analyzed in several different ways to afford a measure of the cardinal parameters (ΔHg, Tg, ΔSg, and ΔCp) that describe the thermodynamics of folding for HPr. The method of Pace and Laurents (Pace CN, Laurents DV, 1989, Biochemistry 28:2520–2525) was used to estimate ΔCP as was a global analysis of the thermal‐ and urea‐induced unfolding data. Each method used to analyze the data gives a similar value for ΔCp (1,170 Δ 50 cal mol−1 K−1). Despite the high melting temperature for HPr (Tg = 73.5 °C), the maximum stability of the protein, which occurs at 26 °C, is quite modest (AGs = 4.2 kcal mol−1). In the presence of moderate concentrations of urea, HPr exhibits cold denaturation, and thus a complete stability curve for HPr, including a measure of ΔCp, can be achieved using the method of Chen and Schellman (Chen B, Schellman JA, 1989, Biochemistry 28:685–691). A comparison of the different methods for the analysis of solvent denaturation curves is provided and the effects of urea on the thermal stability of this small globular protein are discussed. The methods presented will be of general utility in the characterization of the stability curve for many small proteins.
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影响因子：
2.9
作者：
BOLEN, DW;SANTORO, MM
通讯作者：
SANTORO, MM
影响因子：
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作者：
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影响因子：
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作者：
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影响因子：
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作者：
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影响因子：
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作者：
ALEXANDER, P;FAHNESTOCK, S;BRYAN, P
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