The conformation of oxytocin in dimethylsulfoxide as revealed by carbon-13 spin-lattice relaxation times

Information was obtained on rates of overall molecular reorientation and segmental motion of amino acid sidechains of oxytocin in dimethylsulfoxide by determination of spin-lattice relaxation times (T₁) at 25 MHz for carbon-13 in natural abundance in the hormone. The T₁ values of the α-carbons of amino acid residues located in the 20-membered ring of oxytocin are all about 50 msec. The overall correlation time for the hormone backbone was estimated to be 8.8 × 10^?10 sec. The sidechains of Tyr, Ile and Gln undergo segmental motion with respect to the backbone of the ring. The T₁ value of the α-carbon of the Leu residue is greater than for any α-carbon in the ring, indicating an increased mobility of the backbone of the C-terminal acyclic peptide as compared to the ring. The β- and γ-carbons of the Pro residue undergo an exo-endo interconversion with regard to the plane formed by α-carbon, δ-carbon and N atom of the Pro pyrollidine ring. These data are discussed in light of results from other experimental and theoretical studies, including carbon-13 spin-lattice relaxation times for oxytocin in aqueous solution.     

Conformational flexibility of angiotensin II. A carbon-13 spin-lattice relaxation study.

Carbon-13 spin-lattice relaxation times (T1) have been determined for the carbon in the octapeptide hormone [5-isoleucine]-angiotensin II in aqueous solution. Two possible models for molecular motion are considered: isotropic overall motion of the hormone with internal motion of some residues and anisotropic overall molecular motion. The data are interpreted in detail using the former model. The alpha carbons of the peptide backbone are all equally restricted in their motion. The correlation time for overall molecular reorientation, calculated from an everage T1 value of 95 msec for the alpha carbons in the peptide backbone, is ca. 5 times 10-10 sec. The carbons in the side chains are more mobile than those in the peptide backbone, with the exception of the side chain of the Tyr residue which does not undergo rapid segmental motion. We propose that [5-isoleucine]-angiotensin II has a restricted backbone conformation and that the alpha carbons of the N- and C-terminal residues are constrained to nearly the same extent as the remaining alpha carbons in the peptide backbone. Chemical shift data indicate that the Pro residue adopts the trans conformation about the His-Pro bond and that the imidazole ring of His has a strong preference for the N-tau -H tautomer.     

Conformational flexibility of luteinizing hormone-releasing hormone in aqueous solution. A carbon-13 spin-lattice relaxation time study.

The carbon-13 nuclear magnetic resonance (13C NMR) spectra of luteinizing hormone-releasing hormone (LH-RH) and lower homologous peptides have been assigned in aqueous solutions at various pH values. 13C spin-lattice relaxation times (T1) have been measured for all proton-bearing carbons at 25.2 and 67.9 MHz. From the T1 data the rates of overall molecular motion and intramolecular motion of side chains have been estimated. LH-RH is a flexible molecule in solution, having segmental motion along the backbone as well as in the nonaromatic side chains.     

Intramolecular microdynamical and conformational parameters of peptides from 1H and 13C NMR spin-lattice relaxation. Tetragastrin.

Measurements of 1H and 13C spin-lattice relaxation and 13C nuclear Overhauser enhancement factors are reported for dimethyl sulfoxide solutions of tetragastrin, a pharmacologically active tetrapeptide. The use of the dipolar formalism for predicting 1H and quaternary 13C relaxation rates is discussed. Furthermore, the prospect is opened for the use of quaternary 13C and 1H relaxation times to obtain information on the peptide torsion angles phi, psi, and chi in a way supplementing NMR coupling constant methods presently in use.     

Cyclic peptides. XVIII. 13C spin-lattice relaxation times of (X-pro-Y)2 cyclic hexapeptides

¹³C spin-lattice relaxation times (T₁'s) of four cyclic hexapeptides of sequence, (X-L -Pro-Y)₂, are reported. The T₁'s of the protonated carbons, which undergo dipolar relaxation, are interpreted qualitatively in terms of the overall tumbling motion of the molecule and in terms of internal motion. It is found that three of the cyclic hexapeptides, those which adopt all-trans β-conformers, tumble isotropically and appear to lack internal motion in the peptide backbone. The method of Torchia and Lyerla was applied to these compounds in order to compare the mobility of the proline rings. The results show that the sequence and particular type of β-turn present affect the internal motion of the Pro ring. Data on a fourth cyclic hexapeptide, which occurs in a conformation with two-cis X-Pro bonds, suggests that internal motion of the backbone contributes an additional frequency component to the motion of the Y residue α-carbons. A consideration of the mobility of the proline rings in the conformer with two-cis peptide bonds revealed that they are significantly more rigid in the two-cis structure than in the all-trans.     

Carbon-13 spin-lattice relaxation studies of intramolecular motion in lysine and a series of oligolysines

¹³C Spin-lattice relaxation times T₁ for individual carbon nuclei have been measured in a series of oligo-l-lysines, as well as lysine and glycine monomers. Anomalous behavior of profiles of T₁ versus pD occurs for lysine and glycine; the T₁ values of the C_α and CO groups are maximal at pH values corresponding to zwitterionic structures. This is interpreted in terms of the hindered intramolecular rotation around the carbonyl-C_α bond at acidic and basic pD values. Lysine monomer manifests a much less pronounced increase in T₁ values from the α- to the ?-carbon than does lysine in an oligomer or polymer. The rate of reorientation of the C_α and C_β carbons of N-terminal groups are faster than those of the central and C-terminal residues, especially at pD greater than 10 for tri-l-lysine hydrochloride and penta-l-lysine acetate. This is interpreted in terms of interaction between the ?-amino groups and the negatively charged carboxyl groups at pD < 10. Segmental motion is shown to make a significant contribution to relaxation of side-chain carbons, making them less sensitive to molecular size than the carbonyl carbons.     

Hemoglobin aggregation in oxygenated sickle cells studies by carbon-13 rotating frame spin-lattice relaxation in the presence of an off-resonance radiofrequency field

Evidence is presented which shows that hemoglobin S in sickle cells has a tendency to aggregate even in the oxygenated state. The basis for that conclusion is derived from ¹³C-nmr rotating-frame spin–lattice relaxation studies in the presence of an off-resonance radiofrequency field in which the carbonyl resonances of hemoglobins in erythrocytes are examined. The experiments indicate that the rotational correlation time of hemoglobin S in oxygenated sickle cells at 38°C is 130 nsec compared to a value of 95 nsec for hemoglobin A in normal erythrocytes at the same temperature and the same mean cell hemoglobin content.     

ESE relaxation measurements in photosystem II. The influence of the reaction center non-heme iron on the spin-lattice relaxation of Tyr D.

The spin-lattice relaxation of the tyrosine radical D. in Photosystem II particles was studied at 4.2 K in samples in which flash-induced oscillations of the oxidation state of the Mn-cluster of the oxygen evolving system were abolished by addition of ANT2P, leaving Fe2+/Fe3+ oscillations intact. Samples subjected to 0, 1 or 2 light-flashes all showed the same relaxation kinetics. No period-2 oscillation in the spin-lattice relaxation corresponding to the Fe2+/Fe(3+)-oscillation was observed. Thus the T1-oscillations of D. as a function of flash number in untreated samples are solely caused by the charge-oscillations of the Mn-cluster (1989, Biochim, Biophys. Acta 973, 428-442).     

The spin-lattice relaxation times of water associated with early post mortem changes in skeletal muscle.

We studied the spin-echo signal of muscle water in a large time domain and found that the motion of the nuclear magnetic moment of tissue water cannot be characterized by a single spin-lattice relaxation time (T1). The relaxation time T1B, which is the T1 characterized by those protons with a slower relaxation rate, is influenced by the early post mortem changes in skeletal muscle. T1B increased with time after the tissue was taken from the animal and reached a maximum at 3 h. However, the weighted average of T1 of all water protons (T1A) did not change throughout the time course of the experiments.     

Systematic investigation of degradation effects on spin-lattice relaxation times in mouse-liver by low resolution NMR

In spite of numerous work on in vitro proton relaxation time investigations of biological tissue, many questions still remain open. In this study we focused on spin-lattice (T1) relaxation time measurements of mouse liver tissue in order to estimate the time-after-excision effects. The post mortem behaviour of excised tissue was measured up to four hours in intervals of about nine minutes. The time course of liver T1 was determined for different temperatures (4 degrees-40 degrees C) for female mice and the effect of starvation (up to 48 hours) on the time course of T1 was investigated for male and female mice at 37 degrees C. We obtained significant differences in liver T1 time course after excision due to different physiological states like sex, starvation and circadian rhythm.