Department of Medicine, Thoracic Diseases Research Unit, Mayo Clinic, Rochester, Minnesota, USA.
Department of Biochemistry, Thoracic Diseases Research Unit, Mayo Clinic, Rochester, Minnesota, USA.
Antimicrob Agents Chemother. 2024 Mar 6;68(3):e0075623. doi: 10.1128/aac.00756-23. Epub 2024 Jan 23.
cyst life forms contain abundant β-glucan carbohydrates, synthesized using β-1,3 and β-1,6 glucan synthase enzymes and the donor uridine diphosphate (UDP)-glucose. In yeast, phosphoglucomutase (PGM) plays a crucial role in carbohydrate metabolism by interconverting glucose 1-phosphate and glucose 6-phosphate, a vital step in UDP pools for β-glucan cell wall formation. This pathway has not yet been defined in . Herein, we surveyed the and genomes, which predicted a homolog of the major PGM enzyme. Furthermore, we show that PjPgm2p and PmPgm2p function similarly to the yeast counterpart. When both homologs are heterologously expressed in cells, both genes can restore growth and sedimentation rates to wild-type levels. Additionally, we demonstrate that yeast cell lysates expressing the two transcripts individually can restore PGM activities significantly altered in the yeast strain. The addition of lithium, a competitive inhibitor of yeast PGM activity, significantly reduces PGM activity. Next, we tested the effects of lithium on viability and found the compound displays significant anti- activity. Finally, we demonstrate that a para-aryl derivative (ISFP10) with known inhibitory activity against the PGM protein and exhibiting 50-fold selectivity over the human PGM enzyme homolog can also significantly reduce Pmpgm2 activity . Collectively, our data genetically and functionally validate phosphoglucomutases in both and and suggest the potential of this protein as a selective therapeutic target for individuals with pneumonia.
囊泡藻生活形式含有丰富的β-葡聚糖碳水化合物，这些碳水化合物是利用β-1,3 和 β-1,6 葡聚糖合酶以及供体尿苷二磷酸（UDP）-葡萄糖合成的。在酵母中，磷酸葡萄糖变位酶（PGM）通过互变葡萄糖 1-磷酸和葡萄糖 6-磷酸在碳水化合物代谢中起着至关重要的作用，这是 UDP 池用于β-葡聚糖细胞壁形成的重要步骤。在 中，这条途径尚未被定义。在此，我们调查了 和 基因组，预测了 主要 PGM 酶的同源物。此外，我们还表明 PjPgm2p 和 PmPgm2p 与酵母对应物的功能相似。当这两个 同源物在 细胞中异源表达时，两个基因都可以恢复到野生型水平的生长和沉降速率。此外，我们证明酵母 细胞裂解物单独表达这两种 转录本可以显著恢复酵母 菌株中 PGM 活性的改变。添加锂，一种酵母 PGM 活性的竞争性抑制剂，可显著降低 PGM 活性。接下来，我们测试了锂对 活力的影响，发现该化合物对 表现出显著的抗 活性。最后，我们证明了一种具有已知抑制 PGM 蛋白活性的对-芳基衍生物（ISFP10），其对人类 PGM 酶同源物的选择性是 50 倍，也可以显著降低 Pmpgm2 活性。总的来说，我们的数据在 和 中从遗传和功能上验证了磷酸葡萄糖变位酶，并表明该蛋白作为 肺炎患者的选择性治疗靶点具有潜在的可能性。
