A study on the complexes between human erythrocyte enzymes participating in the conversions of 1,3-diphosphoglycerate.
A study on the complexes between human erythrocyte enzymes participating in the conversions of 1,3-diphosphoglycerate.
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参与 1,3-二磷酸甘油酸转化的人红细胞酶之间的复合物的研究。
DOI：
10.1006/abbi.1997.0222
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发表时间：
1997
影响因子：
3.9
通讯作者：
V. Muronetz
中科院分区：
文献类型：
--
作者：
Ksenia V. Fokina;Maria B. Dainyak;N. Nagradova;V. Muronetz
研究了D-甘油醛-3-磷酸脱氢酶（GAPDH）催化人红细胞合成1，3-二磷酸甘油酸的能力，并与以该代谢产物为底物的酶（3-磷酸甘油酸激酶（3-PGK）或2，3-二磷酸甘油酸激酶（2，3-DPGM））形成复合物。结果发现，高活性的2，3-DPGM可以从人红细胞溶血产物中提取，在pH 6.5下与吸附在琼脂糖结合的抗GAPDH抗体上的GAPDH形成复合物，摩尔比为每个GAPDH亚基一个2，3-GPGM亚基。然而，在pH 8.0下未检测到络合。GAPDH和3-PGK之间的相互作用正好相反，在pH 8.0时可以观察到。在pH 7.4下进行的实验中，检测到GAPDH x 2，3-DPGM和GAhGH x 3-PGK复合物。当通过可溶性2，3-DPGM或3-PGK进行共价结合至CNBr活化的琼脂糖凝胶的GAPDH的滴定时，用纯化的酶制剂测定的GAPDH x 2，3-DPGM和GAPDH x 3-PGK酶对的复合物的Kd值在2.40-2.48 μ M的范围内。然而，如果在滴定实验中使用吸附在与琼脂糖凝胶共价结合的特异性抗体上的GAPDH，则发现GAPDH × 2，3-DPGM复合物的Kd为0.54 μ M，GAPDH × 3-PGK复合物的Kd为0.49 μ M。如果在pH 6.5下进行孵育，则在葡萄糖存在下孵育红细胞1小时后测定的2，3-二磷酸甘油酸浓度增加1.5倍，但在pH 8.0下孵育后未发生变化。另一方面，在pH 8.0下孵育后的3-磷酸甘油酸浓度是在pH 6.5下孵育后发现的浓度的两倍。结果解释的假设，GAPDH和2，3-DPGM之间或GAPDH和3-PGK之间的特定蛋白质-蛋白质相互作用可能发挥作用，在确定的命运GAPDH催化的反应中产生的1，3-二磷酸甘油酸。
The ability of D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzing the reaction of 1,3-diphosphoglycerate synthesis in human erythrocytes to form complexes with enzymes which use this metabolite as substrate (3-phosphoglycerate kinase (3-PGK) or 2,3-diphosphoglycerate mutase (2,3-DPGM)) was studied. It was found that highly active 2,3-DPGM can be extracted from human erythrocyte hemolysates in a complex with GAPDH adsorbed on Sepharose-bound anti-GAPDH antibodies at pH 6.5, the molar ratio being one 2,3-GPGM subunit per subunit of GAPDH. No complexation was, however, detected at pH 8.0. The opposite was true for the interaction between GAPDH and 3-PGK, which could be observed at pH 8.0. In experiments carried out at pH 7.4, both GAPDH x 2,3-DPGM and GAPGH x 3-PGK complexes were detected. The Kd values of the complexes determined with purified enzyme preparations were in the range 2.40-2.48 microM for both the GAPDH x 2,3-DPGM and GAPGH x 3-PGK enzyme pairs, when titrations of GAPDH covalently bound to CNBr-activated Sepharose were performed by the soluble 2,3-DPGM or 3-PGK. If, however, GAPDH adsorbed on the specific antibodies covalently bound to Sepharose was used in the titration experiments, the Kd for the GAPDH x 2,3-DPGM complex was found to be 0.54 microM, and the Kd for the GAPDH x 3-PGK complex was 0.49 microM. The concentration of 2,3-diphosphoglycerate determined after 1 h of incubation of erythrocytes in the presence of glucose was found to increase 1.5-fold if the incubation was carried out at pH 6.5, but did not change upon incubation at pH 8.0. On the other hand, the concentration of 3-phosphoglycerate after incubation at pH 8.0 was twice as large as that found after incubation at pH 6.5. The results are interpreted on the hypothesis that specific protein-protein interactions between GAPDH and 2,3-DPGM or between GAPDH and 3-PGK may play a role in determining the fate of 1,3-diphosphoglycerate produced in the GAPDH-catalyzed reaction.
DOI：
10.1016/s0021-9258(20)64292-2
发表时间：
1991-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Harrison;P. Rathinavelu;P. Arese;R. Geahlen;P. Low
通讯作者：
M. Harrison;P. Rathinavelu;P. Arese;R. Geahlen;P. Low
2,3-二磷酸甘油酸和细胞内 pH 作为脱氧血红蛋白 S 生理溶解度的相互依赖的决定因素。
影响因子：
20.3
作者：
Poillon,WN;Kim,BC
通讯作者：
Kim,BC