Regulation of glycolysis by a novel brain-type isozyme of fructose 6-phosphate 2kinase/fructose 2,6-bisphosphatase

果糖6-磷酸2激酶/果糖2,6-二磷酸酶的新型脑型同工酶对糖酵解的调节

• 批准号: 07670157
• 负责人: WATANABE Fusao
• 金额: $ 1.34万
• 依托单位: OSAKA MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670157/
• 关键词: glycolysis; fructose 2,6 bisphospate; brain; fructose 6-phosphate 2-kinase; fructose 2,6-bisphosphatase; alternative splicing; ホスホフルクトキナーゼ; フルクトース2.6二燐酸合成酵素; 分子生物学

Fructose 6-phospate, 2-kinase/fructose 2,6-bisphosphatase catalyzes the synthesis and degradation of fructose 2,6-bisphosphate, which is the most potent activator of glycolysis. We have shown previouslt the occurrence of a partial cDNA (RB7) encoding the catalytic core domain of a novel brain type isozyme. To elucidate the full-length sequence of RB7 cDNA,we have carried out cDNA cloning using 3'- and 5'- rapid amplification cDNA ends method and have isolated eight isoforms from rat brain. The cDNA sequences encoding the 5'-untranslated region, the amino-terminal domain, and the catalytic core domain were identical among all th isoforms. However, heterogeneity of the carboxyl-terminus was found by sequence analysis. This heterogeneity was shown not to have resulted from transcription of multiple genes, as Southern blot and genomic sequence analysis revealed that the gene was a single copy in the rat genome. It is likely that these transcripts represent splice variants of the gene. High-level expression of the gene was also observed northern blot analysis in skeletal muscle. However, the pattern of alternative splicing was different from that of brain, and only four isoforms were detected in skeletal muscle.

6-磷酸果糖，2-激酶/2，6-二磷酸果糖酶催化2，6-二磷酸果糖的合成和降解，2，6-二磷酸是糖酵解最有效的激活剂。我们已经证明了编码一种新的脑型同工酶的催化核心区的部分cDNA(RB7)的出现。为了阐明RB7基因的全长序列，我们用3‘-和5’-快速扩增cDNAEnds方法进行了克隆，并从大鼠脑中分离出了8个亚型。编码5‘-非翻译区、氨基末端结构域和催化核心区的基因序列在所有TH亚型中都是相同的。然而，序列分析发现，该基因的羧基末端具有异质性。这种异质性被证明不是多个基因转录的结果，因为Southern杂交和基因组序列分析表明该基因在大鼠基因组中是单拷贝的。这些转录本很可能代表该基因的剪接变体。Northern印迹分析也观察到该基因在骨骼肌中的高表达。然而，选择性剪接的模式与脑不同，在骨骼肌中只检测到四种异构体。

项目成果

WATANABE,F.et al: "Navel Isoforms of Ret Brain Fructose phosphate,2-Kinase 1 Fructose 2,6-bisphosphatase Are Generated by Tissue-Specific Alteratiue Splicing" Journal of Neurochemistry. (in press). (1997)

WATANABE,F.等人：“Ret 脑果糖磷酸，2-激酶 1 果糖 2,6-二磷酸酶的脐同工型是由组织特异性改变剪接生成的”神经化学杂志。

渡邊 房男: "フルクトース2、6二燐酸合成酵素の新しい脳型アイソザイム" 生化学. 67. 864 (1995)

Fusao Watanabe：“果糖 2,6 二磷酸合酶的新脑型同工酶”生物化学 67. 864 (1995)。

WATANABE, F, et al: "Novel Isoforms of rat brain fructose 6-phosphate, 2-kinasel frutose 2,6-bisphosphatase are generated by tissue-specific alterative splicing" J. Neurochem.69(In press). (1997)

WATANABE, F 等人：“大鼠脑果糖 6-磷酸、2-kinasel 果糖 2,6-二磷酸酶的新型异构体是通过组织特异性可变剪接产生的”J. Neurochem.69（正在印刷中）。

Watanabe, F.et al.: "Enzymatic and fluorescence studies of four single-tryptophan mutants of rat testis fructose 6-phosphate, 2-kinase : fructose 2,6-bisphosphatase." Protein Sci.5. 904-913 (1996)

Watanabe, F. 等人：“大鼠睾丸果糖 6-磷酸，2-激酶：果糖 2,6-双磷酸酶的四种单色氨酸突变体的酶学和荧光研究。”

WATANABE, F. et al: "Enzymatic and fluorescence studies of four single-tryptophan mutants of rat testis fructose 6-phosphate, 2-kinase:fructose 2,6-bishosphatase" Protein Sci.5. 904-913 (1996)

WATANABE, F. 等人：“大鼠睾丸果糖 6-磷酸、2-激酶：果糖 2,6-二磷酸酶的四种单色氨酸突变体的酶学和荧光研究”Protein Sci.5。