title: Selective protein-protein interactions direct channeling of intermediates between polyketide synthase modules.
abstract: Polyketide synthases (PKSs) have represented fertile targets for rational manipulation via protein engineering ever since their modular architecture was first recognized. However, the mechanistic principles by which biosynthetic intermediates are sequentially channeled between modules remain poorly understood. Here we demonstrate the importance of complementarity in a remarkably simple, repetitive structural motif within these megasynthases that has been implicated to affect intermodular chain transfer [Gokhale, R. S., et al. (1999) Science 284, 482]. The C- and N-terminal ends of adjacent PKS polypeptides are capped by short peptides of 20-40 residues. Mismatched sequences abolish intermodular chain transfer without affecting the activity of individual modules, whereas matched sequences can facilitate the channeling of intermediates between ordinarily nonconsecutive modules. Thus, in addition to substrate-PKS interactions and domain-domain interactions, these short interpolypeptide sequences represent a third determinant of selective chain transfer that must be taken into consideration in the protein engineering of PKSs. Preliminary biophysical studies on synthetic peptide mimics of these linkers suggest that they may adopt coiled-coil conformations.
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, California 94305, USA.
authors.forename: S Y
authors.initials: SY
authors.lastname: Tsuji
authors.forename: D E
authors.initials: DE
authors.lastname: Cane
authors.forename: C
authors.initials: C
authors.lastname: Khosla
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D003512:Cyclohexanones / Q000037:antagonists & inhibitors; D004187:Disaccharides / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004917:Erythromycin / Q000031:analogs & derivatives / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology; D015202:Protein Engineering; D017433:Protein Structure, Secondary / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013869:Thiolester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D003512:Cyclohexanones; D004187:Disaccharides; D009097:Multienzyme Complexes; D010446:Peptide Fragments; D011506:Proteins; D011993:Recombinant Fusion Proteins; C413265:tetraketide; C034277:6-deoxyerythronolide B; D004917:Erythromycin; D013869:Thiolester Hydrolases