title: Analysis of the ionic interaction between the hydrophobin RodA and two cutinases of Aspergillus nidulans obtained via an Aspergillus oryzae expression system.
abstract: Hydrophobins are amphipathic secretory proteins with eight conserved cysteine residues and are ubiquitous among filamentous fungi. In the fungus Aspergillus 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 fungus Aspergillus nidulans, which was obtained via an A. oryzae expression 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 recombinant A. nidulans RodA interacted via ionic interactions with two recombinant A. nidulans cutinases. Together, these results imply that ionic interactions between hydrophobins and cutinases may be common among aspergilli and other filamentous fungi.
authors.affiliation: Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, 980-0845, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Microbial Genomics Laboratory, New Industry Creation Hatchery Center, Tohoku University, Sendai, Miyagi, 980-8579, Japan.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Nakayama
authors.affiliation: Microbial Genomics Laboratory, New Industry Creation Hatchery Center, Tohoku University, Sendai, Miyagi, 980-8579, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Department of Microbial Resources, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, 980-0845, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Nanatani
authors.affiliation: Department of Applied Molecular Biology and Biochemistry, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, 183-8509, Japan.
authors.forename: Youhei
authors.initials: Y
authors.lastname: Yamagata
authors.affiliation: Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, 980-0845, Japan. kabe@biochem.tohoku.ac.jp.|Microbial Genomics Laboratory, New Industry Creation Hatchery Center, Tohoku University, Sendai, Miyagi, 980-8579, Japan. kabe@biochem.tohoku.ac.jp.|Department of Microbial Resources, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, 980-0845, Japan. kabe@biochem.tohoku.ac.jp.
authors.forename: Keietsu
authors.initials: K
authors.lastname: Abe
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics* / Q000378:metabolism; D001236:Aspergillus oryzae / Q000235:genetics* / Q000378:metabolism; D063566:Biodegradable Plastics / Q000737:chemistry / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017124:Conserved Sequence; D004950:Esterases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D010802:Phylogeny; D011108:Polymers / Q000737:chemistry / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D055672:Static Electricity
chemical_list: D063566:Biodegradable Plastics; D005656:Fungal Proteins; D007527:Isoenzymes; D011108:Polymers; D011994:Recombinant Proteins; C574545:polybutylene succinate-co-adipate; D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C019952:cutinase
keywords: Aspergillus; Cutinase; Filamentous fungi; Hydrophobin; Ionic interaction