Induction of secondary structure in the peptide hormone motilin by interaction with phospholipid vesicles.
Motilin is an intestinal peptide hormone that binds to a membrane bound receptor located in the gut tissue. Circular dichroism (CD) was used to study the interaction between either porcine or rabbit motilin or a 1-16 fragment of porcine motilin, with model systems of lipid membranes: sodium dodecyl sulphate (SDS), 1,2-dioleoyl-sn-glycero-3-phosphoglycerol (DOPG) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). The CD measurements show significant induction of secondary structure in both motilins and the fragment when negatively charged vesicles (DOPG) or negatively charged micelles (SDS) were present. In contrast, neutral DOPC vesicles did not induce any change in the secondary structure compared to water, in which a random-like secondary structure dominates. The induced secondary structure in the presence of DOPG vesicles is very close to that induced by a mixed aqueous solution containing 30% hexafluoroisopropanol, in which previous NMR-studies have resulted in a three-dimensional solution structure of porcine motilin. In both porcine and rabbit motilin the alpha-helix content is about 50%. This is in agreement with the presence of an amphipathic helix in the C-terminal half of motilin interacting with phospholipid membranes. The interaction appears to be mainly electrostatic in nature, and does not induce any significant alterations in the vesicle, as monitored by EPR studies of spin labels located at the fifth carbon atom of the backbone in a stearic acid molecule. In the 1-16 fragment the alpha-helical content induced by DOPG and SDS is only about 20%.
Backlund
B M
BM
Department of Medical Biochemistry and Biophysics, University of Umeå, Sweden.
Wikander
G
G
Peeters
T L
TL
Gräslund
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D008969:Molecular Sequence Data; D009037:Motilin / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011817:Rabbits; D013552:Swine
D010743:Phospholipids; D009037:Motilin