Molecular mobilities of soluble components in the aqueous phase of chromaffin granules.
NMR relaxation times have been used to characterize molecular motion and intermolecular complexes in the aqueous phase of bovine chromaffin granules. Partially relaxed 13C and proton spectra have been obtained at 3 and 25 degrees C. T1 measurements of five protonated carbons on epinephrine (C2, C5, C6, CHOH and NCH3) give a correlation time of 0.15 (10(-9)) s at 25 degrees C for the catechol ring and methine carbon, while the effective correlation time for the NCH3 group is somewhat shorter due to its internal degree of rotational freedom. Resonances of protonated carbons on the soluble protein chromogranin give very similar correlation times: 0.20 (10(-9)) s for the peptide alpha-carbon and 0.2 (10(-9)) s for the methylene sidechain carbons of glutamic acid. The correlation time (tauR) of ATP was not measured directly using 13C T1 data due to the weakness of its spectrum, but its reorientation appears to be substantially slower than that of epinephrine or chromogranin. This conclusion is based on three observations: (1) the qualitative temperature dependence of T1 for H2 and H8 on the adenine ring places tauR for ATP to the right of the T1 minimum, or tauR greater than or equal to 1.0 (10(-9)) s; (2) 13C-resonances of ATP have anomalously low amplitudes compared with epinephrine resonances, a fact that is readily explained only if ATP undergoes substantially slower reorientation; and (3) a comparison of the T1 data of H8 in chromaffin granules and in a dilute aqueous solution, where tauR for ATP can be measured directly indicates that tauR approximately 1.0 (10(-9)) s at 25 degrees C in the granules. The relaxation data are consistent with the concept of a storage complex based on electrostatic interactions between a polyion (chromogranin) and its counterions (ATP and epinephrine), in which ATP cross-links cationic sidechains of the protein.
D000255:Adenosine Triphosphate; D000313:Adrenal Medulla / Q000166:cytology; D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D004837:Epinephrine; D009682:Magnetic Resonance Spectroscopy; D011522:Protons
D011522:Protons; D000255:Adenosine Triphosphate; D004837:Epinephrine