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991.
The conformation of native and denaturedPhaseolus coccineus var. rubronanus lectin was studied by circular dichroism (CD) and correlated to the hemagglutinating activity. The far-UV CD spectrum at 25°C showed a broad, negative band around 223 nm and a positive one at 196 nm. CD data analysis of the lectin indicated a -sheet-rich protein. At high temperatures, the spectrum was blue-shifted with increasing magnitude; these changes correlated well with the loss of the activity. The conformation of lectin betweenpH 2 and 10 remained essentially unchanged. AtpH 13 the CD spectrum resembled that of unordered form with a negative band near 200 nm and the activity was completely lost. The denatured lectin in 6 M guanidine hydrochloride would be renatured upon diluting the denaturant to 0.75 M; the changes in CD spectrum again correlated well with the loss of the activity. The effect of sodium dodecyl sulfate on the lectin was drastic; it sharply increased thea-helix at the expense of the -sheet and reduced the activity; the changes reached a plateau above 20 mM surfactant. 相似文献
992.
Sharon X. Chen Charles C. Hardin Harold E. Swaisgood 《Journal of Protein Chemistry》1993,12(5):613-625
Incubation of -lactoglobulin with immobilized trypsin at 5–10°C results in a time-dependent release of several fragments of the core domain in yields approaching 15%. Digests were fractionated by ion-exchange chromatography with a Mono Q HR5/5 column and analyzed after disulfide reduction by polyacrylamide gel electrophoresis in sodium dodecylsulfate. Three fragments with approximate molecular weights of 13.8, 9.6, and 6.7 kD were identified. The fraction from ion-exchange chromatography yielding the 6.7 kD fraction after disulfide reduction was further characterized because it was most homogeneous and gave the highest yield. The C-terminal cleavage site of the 6.7 kD core fragment appeared to be Lys100 or Lys101 as determined by C-terminal amino acid analysis. The exact masses, after reduction with dithiothreitol, are 6195 and 6926 as determined by laser desorption mass spectrometry, corresponding to residues 48–101 and 41–100. Prior to reduction, -lactoglobulin C-terminal residues 149–162 are connected to these core domain fragments as shown by C-terminal analysis and mass spectrometry. Structural studies indicate that these 7.9 and 8.6 kD core domain fragments released by immobilized trypsin retain much of their native structure. CD spectra indicate the presence of antiparallel -sheet structure similar to the native protein but the -helix is lost. Spectra in the aromatic region indicate the existence of tertiary structure. Moreover, structural transitions in urea are completely reversible as measured by CD spectra, although the extrapolated G
D
H20
and the urea concentration at the transition midpoint are lower than for the native protein. The core domain fragments also display apH-dependent binding to immobilizedtrans-retinal as does intact protein. A single endotherm is obtained for both core domain fragments and native protein upon differential scanning calorimetry, but again, the domain is less stable as indicated by a transition peak maxima of 56.9°C as compared with 81.1°C for native protein.Abbreviations used: CD, circular dichroism; CPG, controlled pore glass; DSC, differential scanning calorimetry; DTT, dithiothreitol; FPLC, fast flow liquid chromatography; HPLC, high-performance liquid chromatography; PITC, phenylisothiocyanate; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TEA, triethylamine; UV, ultraviolet. 相似文献
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996.
本文对蛋白质中二硫键附近的残基进行了计算机统计分析,结果发现平行和反平行残基间存在着特异的配对规律。这种残基间的相互作用或识别,可能与蛋白质折叠过程中正确地形成二硫键有关。该结果有助于蛋白质工程设计。 相似文献
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998.
本研究在改进后短程序基础上,对氨基酸分离柱进行了改进。改进后的分离柱长为10cm。比原来20cm长柱分离3—MH的时间缩短了近1/2。实验所得的(回收率为97.59%,分离度0.89±0.02。变异系数1.17)这些指标较国外用其它方法所得的结果有良好的相关性。多次测定结果说明长柱与短柱比较无明显差异。证明了短柱对3—MH含量无影响。这一改进所建立的方法大大地缩短了样品的分析时间,节约了大量进口试剂,开展这方面的工作将有利益提高严重烧伤、创伤后蛋白质代谢和营养学等方面的研究水平。 相似文献
999.
本文报道在我国广西隆林壮族中发现一个罕見的HbQ复合α,β地中海贫血家系。先证者女,18岁,贫血面容,肝脾肿大。化学结构分析确证本Hb变异体为HbQ Thailand[α74(EF3)Asp→His]。血红蛋白组成以及α和β珠蛋白基因分析结果表明,先证者的珠蛋白基因型为-α~Q/-α~T复合β°/β°(IVSI-1G→T/Codon17A→T);先证者父的基因型为-‘α~Q/-复合β~O/β~A(IVSI-1G→T/β~A);先证母的基因型为-α~T/αα复合β~O/β~A(Codon17A→T/β~A)。 相似文献
1000.
以自制的壳聚糖作为载体,用戊二醛作交联剂,优化了固定化条件,研制成壳聚糖固定化木瓜蛋白酶。其活性回收率达到42—53%,操作半衰期达到一个月以上,对热、乙醇以及尿素的稳定性有很大的提高,Km值为0.67×10~2mg/mL,最适温度65—70℃,最适pH8.0,能使啤酒中的蛋白质浓度从56.5mg/L减少到2.7mg/L,可以消除啤酒的低温混浊现象。 相似文献