Non-enzymatic glycosylation (or glycation) and inhibition of the pig heart cytosolic aspartate aminotransferase by glyceraldehyde 3-phosphate.

Journal of enzyme inhibition Pub Date : 2000-01-01
C Fitzgerald, T A Swearengin, G Yeargans, D McWhorter, B Cucchetti, N W Seidler
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Abstract

Glyceraldehyde 3-phosphate (Glyc3P), a glycolytic intermediate, non-enzymatically glycosylated (or glycated) and inhibited the pig heart cytoplasmic aspartate aminotransferase (cAAT). Glyc3P (5.0 mM) decreased cAAT activity by 47% after 1 min at 23 degrees C. cAAT activity remained unchanged after a 24h incubation with either glucose 6-phosphate (5.0 mM) or ribose 5-phosphate (5.0 mM). Increasing the incubation pH from 6.4 to 7.8 or the incubation temperature from 23 degrees C to 50 degrees C enhanced Glyc3P's inhibitory effect on cAAT activity. Glyc3P (250-500 microM) decreased the thermal stability of cAAT as evidenced by lowering the Tm or temperature that caused a 50% irreversible loss of cAAT activity (69 degrees C, control; 58.5 degrees C, 500 microM Glyc3P). Glyc3P decreased cAAT amino group content and increased glycation products, which were measured by adduct formation, fluorescence and protein crosslinking.

3-磷酸甘油醛对猪心脏细胞质天冬氨酸转氨酶的非酶糖基化和抑制作用。
甘油醛3-磷酸(Glyc3P)是一种糖酵解中间体,非酶性糖基化(或糖基化)并抑制猪心脏细胞质天冬氨酸转氨酶(cAAT)。Glyc3P (5.0 mM)在23℃作用1分钟后使cAAT活性降低47%,cAAT活性在6-磷酸葡萄糖(5.0 mM)或5-磷酸核糖(5.0 mM)作用24小时后保持不变。将培养pH从6.4增加到7.8或培养温度从23℃增加到50℃,Glyc3P对cAAT活性的抑制作用增强。Glyc3P(250-500微米)降低了cAAT的热稳定性,降低了Tm或温度,导致cAAT活性不可逆损失50%(69℃,对照;58.5℃,500 microM Glyc3P)。通过加合物形成、荧光和蛋白质交联检测,Glyc3P降低了cAAT氨基含量,增加了糖基化产物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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