Alter B P, Goff S C, Klonowski T J, Garrick M D
Prep Biochem. 1980;10(5):633-44. doi: 10.1080/00327488008061758.
Polyacrylamide gel electrophoresis in urea and Triton X-100 of a hemolysate from human fetal red blood cells produces four major protein bands: alpha, beta, and 2 gamma globin chains. We have verified that the latter two are the G gamma and A gamma globin chains which have respectively glycine or alanine at position 136. After incorporation of either [3H] alanine or [3H] glycine into newly synthesized globin each gamma chain was isolated by preparative electrophoresis. The chains were cleaved with cyanogen bromide at methionines 55 and 133, then subjected to automated sequencing, and the residues from each sequencer turn counted. Glycine incorporation was detected for the third turn (position 136) of the G gamma chain and alanine for the A gamma. Substantial metabolic conversion of [3H] glycine to serine and proline was also noted.
在尿素和 Triton X - 100 存在的条件下，对人胎儿红细胞溶血产物进行聚丙烯酰胺凝胶电泳，可产生四条主要蛋白带：α、β和两条γ珠蛋白链。我们已经证实，后两条是在第 136 位分别含有甘氨酸或丙氨酸的 Gγ和 Aγ珠蛋白链。在将[³H]丙氨酸或[³H]甘氨酸掺入新合成的珠蛋白后，通过制备性电泳分离出每条γ链。用溴化氰在甲硫氨酸 55 和 133 处切割这些链，然后进行自动测序，并对每个测序轮次的残基进行计数。在 Gγ链的第三个轮次（第 136 位）检测到甘氨酸掺入，而在 Aγ链检测到丙氨酸掺入。还注意到[³H]甘氨酸大量代谢转化为丝氨酸和脯氨酸。
