Biophysical Characterization of the Unstructured Cytoplasmic Domain of the Human Neuronal Adhesion Protein Neuroligin 3

Gorde:

Xehetasun bibliografikoak
Argitaratua izan da:	Biophysical Journal vol. 95, no. 4 (Aug 15, 2008), p. 1928-1944
Egile nagusia:	Paz, Aviv
Beste egile batzuk:	Zeev-Ben-Mordehai, Tzviya, Lundqvist, Martin, Sherman, Eilon, Mylonas, Efstratios, Weiner, Lev, Haran, Gilad, Svergun, Dmitri I, Mulder, Frans A A, Sussman, Joel L, Silman, Israel
Argitaratua:	Biophysical Society
Gaiak:	Biophysics > methods Cell Adhesion Molecules > chemistry Cell Adhesion Molecules, Neuronal Computer Simulation Cytoplasm > chemistry Humans Membrane Proteins > chemistry Membrane Proteins > ultrastructure Models, Chemical Models, Molecular Nerve Tissue Proteins > chemistry Nerve Tissue Proteins > ultrastructure Protein Conformation Protein Structure, Tertiary Proteins Molecules Mutation Nervous system Nuclear magnetic resonance > NMR E coli Cholinesterase Adhesion Environmental
Sarrera elektronikoa:	Citation/Abstract Full Text + Graphics Full Text - PDF
Etiketak:	Etiketa erantsi Etiketarik gabe, Izan zaitez lehena erregistro honi etiketa jartzen!

Deskribapena
Laburpena:	Cholinesterase-like adhesion molecules (CLAMs) are a family of neuronal cell adhesion molecules with important roles in synaptogenesis, and in maintaining structural and functional integrity of the nervous system. Our earlier study on the cytoplasmic domain of one of these CLAMs, the Drosophila protein, gliotactin, showed that it is intrinsically unstructured in vitro. Bioinformatic analysis suggested that the cytoplasmic domains of other CLAMs are also intrinsically unstructured, even though they bear no sequence homology to each other or to any known protein. In this study, we overexpress and purify the cytoplasmic domain of human neuroligin 3, notwithstanding its high sensitivity to the Escherichia coli endogenous proteases that cause its rapid degradation. Using bioinformatic analysis, sensitivity to proteases, size exclusion chromatography, fluorescence correlation spectroscopy, analytical ultracentrifugation, small angle x-ray scattering, circular dichroism, electron spin resonance, and nuclear magnetic resonance, we show that the cytoplasmic domain of human neuroligin 3 is intrinsically unstructured. However, several of these techniques indicate that it is not fully extended, but becomes significantly more extended under denaturing conditions. [PUBLICATION ABSTRACT] Cholinesterase-like adhesion molecules (CLAMs) are a family of neuronal cell adhesion molecules with important roles in synaptogenesis, and in maintaining structural and functional integrity of the nervous system. Our earlier study on the cytoplasmic domain of one of these CLAMs, the Drosophila protein, gliotactin, showed that it is intrinsically unstructured in vitro. Bioinformatic analysis suggested that the cytoplasmic domains of other CLAMs are also intrinsically unstructured, even though they bear no sequence homology to each other or to any known protein. In this study, we overexpress and purify the cytoplasmic domain of human neuroligin 3, notwithstanding its high sensitivity to the Escherichia coli endogenous proteases that cause its rapid degradation. Using bioinformatic analysis, sensitivity to proteases, size exclusion chromatography, fluorescence correlation spectroscopy, analytical ultracentrifugation, small angle x-ray scattering, circular dichroism, electron spin resonance, and nuclear magnetic resonance, we show that the cytoplasmic domain of human neuroligin 3 is intrinsically unstructured. However, several of these techniques indicate that it is not fully extended, but becomes significantly more extended under denaturing conditions.
ISSN:	0006-3495 1542-0086
Baliabidea:	Science Database