Tim Kirkman, Alice Sketcher, Vinicius de Morais Barroso, Kelly Ishida, Manuela Tosin, Marcio Vinicius Bertacine Dias
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引用次数: 0
摘要
念珠菌性阴道炎已成为一个全球性的健康问题,它通过院内传播而急剧扩散,死亡率很高。由于对氟康唑和两性霉素 B 的广泛耐药性以及对一线药物棘白菌素的耐药性不断增加,目前对念珠菌感染的抗真菌治疗非常有限。因此,迫切需要新的治疗方法来对付这种病原体。二氢叶酸还原酶(DHFR)已被证实是念珠菌的潜在药物靶点,但目前还没有关于念珠菌酶(CauDHFR)结构的报道。本文以近原子分辨率报告了 CauDHFR 的同源酶、全酶以及与嘧霉胺和环胍基(常见的抗喹诺酮类药物)的两种三元复合物的晶体结构。此外,还进行了初步的生化和生物物理检测,以及与多种经典抗真菌剂的抗真菌药敏试验,突出显示了酶抑制率和对酵母生长的抑制作用。这些结构和功能数据可能为针对这一全球性威胁的新药研发活动奠定基础。
Crystal structure of dihydrofolate reductase from the emerging pathogenic fungus Candida auris.
Candida auris has emerged as a global health problem with a dramatic spread by nosocomial transmission and a high mortality rate. Antifungal therapy for C. auris infections is currently limited due to widespread resistance to fluconazole and amphotericin B and increasing resistance to the front-line drug echinocandin. Therefore, new treatments are urgently required to combat this pathogen. Dihydrofolate reductase (DHFR) has been validated as a potential drug target for Candida species, although no structure of the C. auris enzyme (CauDHFR) has been reported. Here, crystal structures of CauDHFR are reported as an apoenzyme, as a holoenzyme and in two ternary complexes with pyrimethamine and cycloguanil, which are common antifolates, at near-atomic resolution. Preliminary biochemical and biophysical assays and antifungal susceptibility testing with a variety of classical antifolates were also performed, highlighting the enzyme-inhibition rates and the inhibition of yeast growth. These structural and functional data might provide the basis for a novel drug-discovery campaign against this global threat.
期刊介绍:
Acta Crystallographica Section D welcomes the submission of articles covering any aspect of structural biology, with a particular emphasis on the structures of biological macromolecules or the methods used to determine them.
Reports on new structures of biological importance may address the smallest macromolecules to the largest complex molecular machines. These structures may have been determined using any structural biology technique including crystallography, NMR, cryoEM and/or other techniques. The key criterion is that such articles must present significant new insights into biological, chemical or medical sciences. The inclusion of complementary data that support the conclusions drawn from the structural studies (such as binding studies, mass spectrometry, enzyme assays, or analysis of mutants or other modified forms of biological macromolecule) is encouraged.
Methods articles may include new approaches to any aspect of biological structure determination or structure analysis but will only be accepted where they focus on new methods that are demonstrated to be of general applicability and importance to structural biology. Articles describing particularly difficult problems in structural biology are also welcomed, if the analysis would provide useful insights to others facing similar problems.