title: Biochemical characterization of human dynamin-like protein 1.
abstract: Human dynamin-like protein 1 (DLP-1) is involved in the fission of mitochondrial outer membranes, a process that helps to maintain mitochondrial morphology and to reduce the accumulation of functional and structural defects in mitochondria. DLP-1 is a ~80 kDa membrane-interacting protein and contains a GTPase domain, a middle domain, a putative PH-like domain and a GTPase effector domain (GED). While the GED has been suggested to be important on protein oligomerization and GTPase activation, functional relationships between the other domains especially the roles of the middle domain in protein activity remains less clear. In this study, we have investigated the biochemical properties of recombinant DLP-1 wild-type and selected mutants, all expressed in Escherichia coli. The middle domain mutants G350D, R365S and ΔPH (lacking the putative PH-like domain) severely impair the GTPase activity, but have no obvious effects on protein tetramerization and liposome-binding properties, suggesting these mutants probably affect protein intra-molecular interactions. Our study also suggested that proper domain-domain interactions are important for DLP-1 GTPase activity.
authors.affiliation: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824-1319, USA.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Gao
authors.forename: R Michael
authors.initials: RM
authors.lastname: Garavito
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D034281:Dynamins; D004789:Enzyme Activation; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000235:genetics; D008928:Mitochondria / Q000737:chemistry; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D020539:Sequence Analysis, Protein
chemical_list: D008081:Liposomes; D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins