A hydrophobic patch in the competence-stimulating Peptide, a pneumococcal competence pheromone, is essential for specificity and biological activity.
Induction of competence for natural genetic transformation in Streptococcus pneumoniae depends on pheromone-mediated cell-cell communication and a signaling pathway consisting of the competence-stimulating peptide (CSP), its membrane-embedded histidine kinase receptor ComD, and the cognate response regulator ComE. Extensive screening of pneumococcal isolates has revealed that two major CSP variants, CSP1 and CSP2, are found in members of this species. Even though the primary structures of CSP1 and CSP2 are about 50% identical, they are highly specific for their respective receptors, ComD1 and ComD2. In the present work, we have investigated the structural basis of this specificity by determining the three-dimensional structure of CSP1 from nuclear magnetic resonance data and comparing the agonist activity of a number of CSP1/CSP2 hybrid peptides toward the ComD1 and ComD2 receptors. Our results show that upon exposure to membrane-mimicking environments, the 17-amino-acid CSP1 pheromone adopts an amphiphilic alpha-helical configuration stretching from residue 6 to residue 12. Furthermore, the pattern of agonist activity displayed by the various hybrid peptides revealed that hydrophobic amino acids, some of which are situated on the nonpolar side of the alpha-helix, strongly contribute to CSP specificity. Together, these data indicate that the identified alpha-helix is an important structural feature of CSP1 which is essential for effective receptor recognition under natural conditions.
Johnsborg
Ola
O
Department of Chemistry, Biotechnology, and Food Science, Biotechnology Building, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 As, Norway.
Kristiansen
Per Eugen
PE
Blomqvist
Trinelise
T
Håvarstein
Leiv Sigve
LS
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary / Q000235:genetics; D043563:Receptors, Pheromone / Q000378:metabolism*; D013045:Species Specificity; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D043563:Receptors, Pheromone; C423628:competence factor, Streptococcus