Dansylation of tyrosine: hindrance by N-ethylmorpholine and photodegradation of O-dansylated derivatives.
Dansylation of tyrosine: hindrance by N-ethylmorpholine and photodegradation of O-dansylated derivatives.
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酪氨酸的丹磺酰化：N-乙基吗啉的阻碍和 O-丹磺酰化衍生物的光降解。
DOI：
10.1016/0003-2697(77)90684-4
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发表时间：
1977
影响因子：
2.9
通讯作者：
J. Wilson
中科院分区：
文献类型：
--
作者：
P. Felgner;J. Wilson
如果在丹酰化过程中使用N-乙基吗啉，则游离酪氨酸或含有N-末端酪氨酸残基的鼠脑己糖激酶（ATP：d-己糖6-磷酸转移酶：EC 2.7.1.1）的丹酰化产生双丹酰和N-单丹酰衍生物[W. R. Gray（1972）in Methods in Enzymology（Hirs，C. H. W.，和Timasheff，S. N.，编辑），Vol. 25，pp. 121-138，Academic Press，纽约]。如果用NaHCO 3缓冲液（pH 9.5）代替N-乙基吗啉，则几乎只形成二丹磺酰基衍生物，从而消除了由单个N-末端残基形成两种衍生物所导致的不确定性。因此，一个稍微修改的丹磺酰化程序，使用NaHCO 3缓冲液的建议，其成功的应用程序，以六种不同的蛋白质具有先前已知的N-末端氨基酸的修改后的程序的有效性得到了证明。与N-丹磺酰基衍生物相比，酪氨酸的二丹磺酰基和O-单丹磺酰基衍生物是显著的光不稳定的。除非观察到针对不必要的照射的特定预防措施，否则在实验操作期间可能发生二丹酰酪氨酸衍生物的光解降解;二丹酰化化合物的损失和光解产物的形成使对其中预期单个二丹酰衍生物（N-末端残基）的实验的解释复杂化。
Dansylation of free tyrosine or of rat brain hexokinase (ATP:d-hexose 6-phosphotransferase: EC 2.7.1.1), which contains an N-terminal tyrosine residue, yields both the didansyl and N-monodansyl derivatives if N-ethylmorpholine is used in the dansylation procedure [W. R. Gray (1972)in Methods in Enzymology (Hirs, C. H. W., and Timasheff, S. N., eds.), Vol. 25, pp. 121–138, Academic Press, New York]. If the N-ethylmorpholine is replaced by NaHCO3buffer (pH 9.5), the didansyl derivative is formed almost exclusively, and the ambiguity resulting from the formation of two derivatives from a single N-terminal residue is thereby eliminated. Therefore, a slightly modified dansylation procedure, using NaHCO3buffer is recommended; the validity of the modified procedure was demonstrated by its successful application to six different proteins having previously known N-terminal amino acids. The didansyl and O-monodansyl derivatives of tyrosine are remarkably photolabile as compared to the N-dansyl derivatives. Unless specific precautions against unnecessary irradiation are observed, photolytic degradation of the didansyl tyrosine derivatives could occur during experimental manipulations; loss of the didansylated compound and formation of photolysis products complicates the interpretation of experiments in which a single didansyl derivative (of the N-terminal residue) is expected.