NADP-dependent glutamate dehydrogenases in a dimorphic zygomycete Benjaminiella poitrasii: Purification, characterization and their evaluation as an antifungal drug target.,Biochimica et Biophysica Acta (BBA) - General Subjects

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NADP-dependent glutamate dehydrogenases in a dimorphic zygomycete Benjaminiella poitrasii: Purification, characterization and their evaluation as an antifungal drug target.
Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 2.8 ) Pub Date : 2020-08-06 , DOI: 10.1016/j.bbagen.2020.129696
Ejaj K Pathan ₁ , Anand M Kulkarni ₂ , Nallaballe V L Prasanna ₃ , Chepuri V Ramana ₃ , Mukund V Deshpande ₁

Affiliation

Background

It has been reported that the genes coding for NADP-dependent glutamate dehydrogenases (NADP-GDHs) showed a cause-effect relationship with Yeast-Hypha (YH) reversible transition in a zygomycete Benjaminiella poitrasii. As YH transition is significant in human pathogenic fungi for their survival and proliferation in the host, the NADP-GDHs can be explored as antifungal drug targets.

Methods

The yeast-form specific BpNADPGDH I and hyphal-form specific BpNADPGDH II of B. poitrasii were purified by heterologous expression in E. coli BL-21 cells and characterized. The structural analogs of L-glutamate, dimethyl esters of isophthalic acid (DMIP) and its derivatives were designed, synthesized and screened for inhibition of NADP-GDH activity as well as YH transition in B. poitrasii, and also in human pathogenic Candida albicans strains.

Results

The BpNADPGDH I and BpNADPGDH II were found to be homo-hexameric proteins with native molecular mass of 282 kDa and 298 kDa, respectively and subunit molecular weights of 47 kDa and 49 kDa, respectively. Besides the distinct kinetic properties, BpNADPGDH I and BpNADPGDH II were found to be regulated by cAMP-dependent- and Calmodulin (CaM) dependent- protein kinases, respectively. The DMIP compounds showed a more pronounced effect on H-form specific BpNADPGDH II and inhibited YH transition as well as growth in B. poitrasii and C. albicans strains.

Conclusion

The present study will be useful to design and develop antifungal drugs against dimorphic human pathogens using glutamate dehydrogenase as a target.

Significance

Glutamate dehydrogenases can be explored as a target against human pathogenic fungi.

中文翻译：

双态合子菌Benjaminiella poitrasii中NADP依赖的谷氨酸脱氢酶：纯化，表征和其作为抗真菌药物靶标的评估。

背景

据报道，编码NADP依赖性谷氨酸脱氢酶（NADP-GDHs ）的基因在酵母菌Benjaminiella poitrasii中显示出与酵母-菌丝（Y H）可逆转变的因果关系。由于Y H过渡对于人类病原性真菌在宿主中的存活和增殖具有重要意义，因此可以将NADP-GDHs用作抗真菌药物的靶标。

方法

通过在大肠杆菌BL-21细胞中的异源表达，纯化了芽孢杆菌的酵母形式特异性BpNADPGDH I和菌丝形式特异性BpNADPGDH II并进行了表征。设计，合成和筛选了L-谷氨酸，间苯二甲酸二甲酯（DMIP）及其衍生物的结构类似物，以抑制NADP-GDH活性以及Poitrasii以及人致病性白色念珠菌中的Y H转变。株。

结果

发现BpNADPGDH I和BpNADPGDH II是天然分子量分别为282 kDa和298 kDa，亚基分子量分别为47 kDa和49 kDa的同六聚体蛋白。除了独特的动力学特性外，还发现BpNADPGDH I和BpNADPGDH II分别受cAMP依赖性和钙调蛋白（CaM）依赖性蛋白激酶的调节。DMIP化合物对H型特异性BpNADPGDH II表现出更明显的作用，并抑制了Y H过渡以及在Poitrasii和白色念珠菌菌株中的生长。