A single protein research integrated advanced biochemistry laboratory course; spectroscopic determination of tyrosyl side chain pKa

Brian N. Blanchette, Bal Ram Singh
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Abstract

The unique bio-analytical properties of the amino acid tyrosine (Tyr) are the focus of this experiment from the research oriented biochemistry laboratory course at our university. In the present study pKa1, pKa2, and pKa3 values for free Tyr were estimated to be 2.30, 9.40, and 9.97, respectively, when free Tyr was titrated with 1 mM NaOH and 1 mM HCl using a pH meter. Spectrophotometric analysis of the phenolic side chain pKa3 revealed a value of 10.14, which was consistent with the pKas estimated from the pH meter. The results from this experiment will allow students to compare the free Tyr properties with those present in a protein.

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结合高等生物化学实验课程的单蛋白研究酪氨酸侧链pKa的光谱测定
本实验以我校研究型生物化学实验课程中氨基酸酪氨酸(Tyr)独特的生物分析特性为重点。在本研究中,当使用pH计用1mm NaOH和1mm HCl滴定游离Tyr时,pKa1、pKa2和pKa3的值分别为2.30、9.40和9.97。对酚侧链pKa3的分光光度分析显示pKa3的值为10.14,与pH计测得的pKa3值一致。这个实验的结果将允许学生比较游离酪氨酸与蛋白质中存在的特性。
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