Wasylewski Z, Stryjewski W, Waśniowska A, Potempa J, Baran K
Biochim Biophys Acta. 1986 Jun 5;871(2):177-81. doi: 10.1016/0167-4838(86)90171-8.
The removal by EDTA of Ca2+ from the two-tryptophan-containing metalloproteinase isolated from Staphylococcus aureus leads to an increase in its intrinsic fluorescence intensity. Based on acrylamide fluorescence quenching results, analyzed by the non-linear least-squares method, we have shown that this protein molecule undergoes irreversible conformational change upon removal of Ca2+, which include the exposure to the solvent of buried tryptophan residues. Steady-state fluorescence anisotropy measurements indicate that the loss of Ca2+ leads to a significant increase in internal mobility of previously buried tryptophan residues.
从金黄色葡萄球菌中分离得到的含两个色氨酸的金属蛋白酶，用乙二胺四乙酸（EDTA）去除Ca2+后，其固有荧光强度增加。基于用非线性最小二乘法分析的丙烯酰胺荧光猝灭结果，我们表明该蛋白质分子在去除Ca2+后会发生不可逆的构象变化，其中包括埋藏的色氨酸残基暴露于溶剂中。稳态荧光各向异性测量表明，Ca2+的丢失导致先前埋藏的色氨酸残基的内部流动性显著增加。
