Iwase H, Ishii I, Saito T, Ohara S, Hotta K
Department of Biochemistry, School of Medicine, Kitasato University, Kanagawa, Japan.
Anal Biochem. 1988 Sep;173(2):317-20. doi: 10.1016/0003-2697(88)90195-9.
The enzymatic transfer of the sugar portion from UDP-N-acetylgalactosamine to pyridylamino (PA) lacto-N-fucopentaose I (Fuc alpha 1-2Gal beta 1-3GlcNAc beta 1-4Glc-PA) was detected by high-performance liquid chromatography. Separation of the fluorescent product from the fluorescence-labeled acceptor was achieved within 10 min by reversed-phase high-performance liquid chromatography. Rat stomach enzyme activity was detected in the microsomal fraction from antrum but not corpus. Ohara et al. (1986, Comp. Biochem. Physiol. 83B, 273-275) reported that the N-acetylgalactosamine content in antrum mucin was greater than that in corpus mucin and antrum mucin had strong blood group A activity. The prominent asymmetrical distribution of the enzyme detected here well supports these findings. The elution position of the fluorescent product was the same as that of the product formed by the action of type A human serum toward the acceptor. Its hydrolysis by alpha-N-acetylgalactosaminidase yielded the acceptor. It is thus evident that the detected enzyme is the same as that producing the blood group A structure.
通过高效液相色谱法检测了糖部分从UDP-N-乙酰半乳糖胺向吡啶基氨基（PA）乳糖-N-岩藻五糖I（Fucα1-2Galβ1-3GlcNAcβ1-4Glc-PA）的酶促转移。通过反相高效液相色谱法在10分钟内实现了荧光产物与荧光标记受体的分离。在胃窦而非胃体的微粒体部分检测到大鼠胃酶活性。大原等人（1986年，《比较生物化学与生理学》83B，273 - 275）报道，胃窦粘蛋白中的N-乙酰半乳糖胺含量高于胃体粘蛋白，且胃窦粘蛋白具有较强的A血型活性。此处检测到的酶的显著不对称分布很好地支持了这些发现。荧光产物的洗脱位置与A型人血清作用于受体形成的产物相同。其被α-N-乙酰半乳糖胺酶水解产生受体。因此很明显，检测到的酶与产生A血型结构的酶相同。
