title: Ranacyclins, a new family of short cyclic antimicrobial peptides: biological function, mode of action, and parameters involved in target specificity.
abstract: We report on two new cyclic 17-residue peptides that we named ranacyclins E and T, the first isolated from Rana esculenta frog skin secretions and the second discovered by screening a cDNA library from Rana temporaria. Ranacyclins have a loop region that is homologous with that of an 18-mer peptide, pLR, isolated from the skin of the Northern Leopard frog, Rana pipiens, with no reported antimicrobial activity. Here we show that ranacyclins and pLR have antimicrobial and antifungal activity. However, despite the high structural similarity, they differ in their spectrum of activity. The data reveal that ranacyclins and pLR have several properties that differentiate them from most known antimicrobial peptides. These include the following: (i) they adopt a significant portion of random coil structure in the membrane as revealed by ATR-FTIR and CD spectroscopy (50% for ranacyclin T and 70% for both ranacyclin E and pLR); (ii) they bind similarly to both zwitterionic and negatively charged membranes as revealed by using tryptophan fluorescence and surface plasmon resonance (SPR; BIAcore biosensor); (iii) they insert into the hydrophobic core of the membrane and presumably form transmembrane pores without damage to the bacterial wall, as revealed by SPR, ATR-FTIR, and transmission electron microscopy (TEM); and (iv) despite being highly and equally active in permeating bacterial spheroplasts and negatively charged membranes, they differ significantly in their potencies against target cells. Furthermore, a significant fraction of a given secondary structure is not prerequisite for membrane permeation and antimicrobial activity. However, increasing the fraction of a secondary structure and reducing peptide assembly in the membrane make it easier for the peptide to diffuse through the cell wall, which is different for each microorganism, into the cytoplasmic membrane.
authors.affiliation: Istituto Pasteur Fondazione Cenci Bolognetti, Dipartimento di Scienze Biochimiche, Ospedale S. Andrea, and CNR Istituto di Biologia e Patologia Molecolari, Università La Sapienza, 00185 Roma, Italy.
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Mangoni
authors.forename: Niv
authors.initials: N
authors.lastname: Papo
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Mignogna
authors.forename: David
authors.initials: D
authors.lastname: Andreu
authors.forename: Yechiel
authors.initials: Y
authors.lastname: Shai
authors.forename: Donatella
authors.initials: D
authors.lastname: Barra
authors.forename: Maurizio
authors.initials: M
authors.lastname: Simmaco
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000187:drug effects / Q000382:microbiology / Q000502:physiology; D004357:Drug Synergism; D006461:Hemolysis / Q000187:drug effects; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008954:Models, Biological; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D010743:Phospholipids / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D011893:Rana esculenta; D011896:Rana temporaria; D012996:Solutions; D017550:Spectroscopy, Fourier Transform Infrared; D020349:Surface Plasmon Resonance
chemical_list: D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D000935:Antifungal Agents; D010456:Peptides, Cyclic; D010743:Phospholipids; D012996:Solutions; C482660:ranacyclin E; C482661:ranacyclin T