Norén O, Sjöström H, Larsen P
Eur J Biochem. 1983 Jul 15;134(1):71-6. doi: 10.1111/j.1432-1033.1983.tb07532.x.
A procedure for the synthesis of tritiated 4-azidophenylalanine in a radioactively homogeneous form is described. The characteristics of the 4-azidophenylalanine uptake by pig intestinal microvillar vesicles and its interaction with the neutral amino acid transport have been studied. 4-Azidophenylalanine is transported into an osmotically sensitive space and the transport is sodium dependent. It is demonstrated that 4-azidophenylalanine transport is inhibited by L-alanine and L-phenylalanine but not by L-lysine and D-glucose. Likewise 4-azidophenylalanine inhibited the initial transport of L-alanine and L-phenylalanine but the initial transport of L-lysine and D-glucose was unaffected. Treatment of microvillar vesicles with 4-azidophenylalanine in combination with photolysis irreversibly inactivated the L-alanine transport. Photolabelling in the presence of sodium using tritiated 4-azidophenylalanine resulted in labelling of 10 peptides, all found in the fraction insoluble in Triton X-100. Omission of sodium from the reaction mixture or addition of L-alanine did not specifically abolish labelling in any of the labelled components.
本文描述了一种合成放射性均匀形式的氚标记4-叠氮基苯丙氨酸的方法。研究了猪小肠微绒毛小泡对4-叠氮基苯丙氨酸的摄取特性及其与中性氨基酸转运的相互作用。4-叠氮基苯丙氨酸被转运到一个对渗透压敏感的空间，且该转运依赖于钠离子。结果表明，L-丙氨酸和L-苯丙氨酸可抑制4-叠氮基苯丙氨酸的转运，但L-赖氨酸和D-葡萄糖则无此作用。同样，4-叠氮基苯丙氨酸可抑制L-丙氨酸和L-苯丙氨酸的初始转运，但对L-赖氨酸和D-葡萄糖的初始转运无影响。用4-叠氮基苯丙氨酸处理微绒毛小泡并结合光解作用可使L-丙氨酸转运不可逆地失活。在有钠离子存在的情况下，用氚标记的4-叠氮基苯丙氨酸进行光标记，结果标记了10种肽，所有这些肽都存在于不溶于Triton X-100的组分中。从反应混合物中去除钠离子或添加L-丙氨酸并不会特异性地消除任何标记组分中的标记。
