acth-(11-24) and dodecylphosphocholine

Three adrenocorticotropin hormone (ACTH) fragments (1-10, 1-24, and 11-24) have been studied in water and in sodium dodecylsulfate (SDS) and dodecylphosphocholine (DPC) micelles by nuclear magnetic resonance spectroscopy. The trans-cis isomerism at all three proline sites (at positions 12, 19, and 24) was found in the 11-24 segment of the peptide. The population of the cis isomers changes with the environment of the peptide. Specifically, the presence of the DPC micelle does not affect the trans-cis equilibrium in the 11-24 segment from that in water. In contrast, the presence of the SDS micelles decreases the population of the cis isomer at Pro(24), but increases its population at Pro(12) and Pro(19). The effect of SDS micelles on the trans-cis equilibrium at these proline sites was discussed. Intermolecular nuclear Overhauser effect (NOE) correlations between the ACTH peptides and the micelles were observed. These correlations occurred only in the 1-10 segment of the peptides, and the hydrophobic side chains contributed most to the intermolecular NOE. The intermolecular NOE pattern corroborates the suggestion that the 1-10 segment of the ACTH peptides bind to these micelles via a surface-binding mode, with most of the interactions coming from the insertion of the hydrophobic side chains.

Topics: Adrenocorticotropic Hormone; Amino Acid Sequence; Cosyntropin; Isomerism; Micelles; Molecular Sequence Data; Nuclear Magnetic Resonance, Biomolecular; Peptide Fragments; Phosphorylcholine; Proline; Sodium Dodecyl Sulfate

The partition and structure of three adrenocorticotropic hormone peptides ACTH(1-10), ACTH(1-24), and ACTH(11-24) in water and in sodium dodecylsulfate (SDS) and dodecylphosphocholine (DPC) micelles were studied by 2D NMR and NMR gradient diffusion measurements. The diffusion rates, the NH chemical shifts, and the nuclear Overhauser effect patterns provided a coherent picture of binding of these peptides. All three peptides are significantly partitioned in the negatively charged SDS micelles and possess definite secondary structure, as opposed to random structures in water. For ACTH (1-24), the hydrophobic 1-10 segment is partitioned in DPC micelles, but the charged 11-24 segment prefers to remain in the aqueous region. ACTH(11-24) does not bind significantly to the DPC micelles. The binding of the ACTH peptides in these two widely used "membrane mimics" are substantially different from that in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayers obtained by attenuated total reflection infrared spectroscopy and from our preliminary diffusion studies of the same peptides in POPC vesicles. This study showed that, in a given micellar medium, all corresponding segments of these peptides are located in the same membrane environment in the system, regardless of whether these segments exist by themselves or are attached to other segments. This result may contradict the membrane-compartments concept of Schwyzer, which suggests that ACTH(1-10) and ACTH(1-24) are located in different membrane compartments because they have different address segments, and consequently, bind to different receptors. The present results also suggest that the assumption that micelles are good membrane mimics should be carefully examined.

Topics: Adrenocorticotropic Hormone; Biophysical Phenomena; Biophysics; Cosyntropin; Diffusion; In Vitro Techniques; Membranes, Artificial; Micelles; Nuclear Magnetic Resonance, Biomolecular; Peptide Fragments; Phosphorylcholine; Proline; Protein Binding; Protein Structure, Secondary; Sodium Dodecyl Sulfate; Stereoisomerism