Involvement and identification of a tryptophanyl residue at the pyruvate binding site of transcarboxylase.
Involvement and identification of a tryptophanyl residue at the pyruvate binding site of transcarboxylase.
复制标题
转羧酶丙酮酸结合位点色氨酸残基的参与和鉴定。
DOI：
10.1021/bi00416a022
复制
发表时间：
1988
期刊：
影响因子：
2.9
通讯作者：
Wood,HG
中科院分区：
文献类型：
--
作者：
Kumar,GK;Haase,FC;Phillips,NF;Wood,HG
MethodsMethods such as preparation of transcarboxylase and subunits, fluorescence measurements, procedure for trypsinization, and high-performance liquid chromatographyseparation of peptides used in this study were essentially similar to those used by Kumar et al.(1988).Acrylamide Quenching Studies. Fluorescence intensities were determined by using an excitation wavelength of 295 nm and by continuous monitoring of the emission at 340 nm. The final intensity at any given acrylamide concentration was taken as a time-averaged value (= 10). Additions of acrylamide were made from a concentrated (8.0 M) stock solution. Fluorescence was corrected fordilution and inner filter effect (Héléne et al., 1969). A correction for absorption of the acrylamide itself at 295 nm was made according to the method of McClure and Edelman (1967) using a molar extinction coefficient of 0.29 M-1 cm-1 at 295 nm. The details of fluorescence titrations of transcarboxylase and the outer 5S subunit with the substrate pyruvate are described in the figure
