Analysis of the ionic interaction between the hydrophobin RodA and two cutinases of Aspergillus nidulans obtained via an Aspergillus oryzae expression system
Analysis of the ionic interaction between the hydrophobin RodA and two cutinases of Aspergillus nidulans obtained via an Aspergillus oryzae expression system
复制标题
疏水蛋白 RodA 与通过米曲霉表达系统获得的两种构巢曲霉角质酶之间的离子相互作用分析
DOI：
10.1007/s00253-016-7979-5
复制
发表时间：
2017
影响因子：
5
通讯作者：
Keietsu Abe
中科院分区：
文献类型：
--
作者：
Takumi Tanaka;Mayumi Nakayama;Toru Takahashi;Kei Nanatani;Youhei Yamagata;Keietsu Abe
疏水蛋白是具有八个保守半胱氨酸残基的两亲性分泌蛋白，在丝状真菌中普遍存在。在真菌米曲霉中，当唯一可用的碳源是可生物降解的聚酯聚丁二酸己二酸丁二醇酯 (PBSA) 时，疏水蛋白 RolA 和聚酯酶 CutL1 共表达。 RolA 通过附着在颗粒表面、改变其结构并与 CutL1 相互作用使 CutL1 集中在 PBSA 表面来促进 PBSA 的降解。我们之前报道过 RolA 中带正电的残基和 CutL1 中带负电的残基协同参与 RolA 和 CutL1 之间的离子相互作用。我们还报道了模型真菌构巢曲霉的疏水蛋白RodA，它是通过anA获得的。 oryzae表达系统，通过离子相互作用与CutL1相互作用。在本研究中，系统发育和比对分析表明，几个RolA直向同源物的N端区域含有带正电荷的残基，并且CutL1表面上对于RolA-CutL1相互作用至关重要的相应带负电荷的残基在几个CutL1直向同源物中高度保守。 PBSA 微粒降解测定、使用聚四氟乙烯颗粒分散体的下拉测定以及使用石英晶体微天平的动力学分析表明重组A。 nidulansRodA 通过离子相互作用与两个重组 A 相互作用。构巢糖酶。总之，这些结果表明疏水蛋白和角质酶之间的离子相互作用在曲霉和其他丝状真菌中可能很常见。
Hydrophobins are amphipathic secretory proteins with eight conserved cysteine residues and are ubiquitous among filamentous fungi. In the fungusAspergillus oryzae, the hydrophobin RolA and the polyesterase CutL1 are co-expressed when the sole available carbon source is the biodegradable polyester polybutylene succinate-co-adipate (PBSA). RolA promotes the degradation of PBSA by attaching to the particle surface, changing its structure and interacting with CutL1 to concentrate CutL1 on the PBSA surface. We previously reported that positively charged residues in RolA and negatively charged residues in CutL1 are cooperatively involved in the ionic interaction between RolA and CutL1. We also reported that hydrophobin RodA of the model fungusAspergillus nidulans, which was obtained via anA. oryzaeexpression system, interacted via ionic interactions with CutL1. In the present study, phylogenetic and alignment analyses revealed that the N-terminal regions of several RolA orthologs contained positively charged residues and that the corresponding negatively charged residues on the surface of CutL1 that were essential for the RolA–CutL1 interaction were highly conserved in several CutL1 orthologs. A PBSA microparticle degradation assay, a pull-down assay using a dispersion of Teflon particles, and a kinetic analysis using a quartz crystal microbalance revealed that recombinantA. nidulansRodA interacted via ionic interactions with two recombinantA. nidulanscutinases. Together, these results imply that ionic interactions between hydrophobins and cutinases may be common among aspergilli and other filamentous fungi.