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Membrane structure and interactions of human catestatin by multidimensional solution and solid‐state NMR spectroscopy
Author(s) -
Sugawara Masae,
Resende Jarbas M.,
Moraes Cléria Mendonça,
Marquette Arnaud,
Chich JeanFrancois,
MetzBoutigue MarieHélène,
Bechinger Burkhard
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.09-142554
Subject(s) - chemistry , circular dichroism , nuclear magnetic resonance spectroscopy , biophysics , bilayer , solid state nuclear magnetic resonance , micelle , lipid bilayer , membrane , peptide , proton nmr , stereochemistry , biochemistry , crystallography , nuclear magnetic resonance , aqueous solution , biology , physics
Catestatin is a natural peptide of higher organisms including humans, with a wide variety of biological functions involved in catecholamine inhibition, cardiovascular regulation, control of blood pressure, inflammation, and innate immunity. It is derived from the natural processing of chromogranin A, induced in the skin after injury, and produced by chromaffin cells and neutrophils. With neutrophils, the peptide enters the cell by crossing the plasma membrane where it interacts with internal targets to induce calcium influx. Therefore, we investigated the membrane interactions and structure of several catestatin‐derived peptides. Whereas fluorescence dye release experiments are indicative of membrane permeabilization, multidimensional solution NMR and circular dichroism spectroscopies show that catestatin adopts α‐helical conformations between Ser‐6 and Tyr‐12 in the presence of dodecylphosphocholine micelles. Furthermore, proton‐decoupled 15 N solid‐state NMR spectroscopy of sequences labeled with 15 N and reconstituted into oriented lipid bilayers indicates that this domain is aligned in a strongly tilted to inplanar alignment. Proton‐decoupled 31 P NMR spectra of the same samples are indicative of conformational and/or orientational heterogeneity at the level of the lipid bilayer head groups due to the presence of catestatin. The sequence and 3‐dimensional structure of catestatin exhibit homologies with penetratin, which is suggestive that they both enter the cells by related mechanisms to target internal structures.—Sugawara, M., Resende, J. M., Moraes, C. M., Marquette, A., Chich, J.‐F., Metz‐Boutigue, M.‐H., Bechinger, B. Membrane structure and interactions of human catestatin by multidimensional solution and solid‐state NMR spectroscopy. FASEB J. 24, 1737–1146 (2010). www.fasebj.org