title: "CON-CON" assignment strategy for highly flexible intrinsically disordered proteins.
abstract: Intrinsically disordered proteins (IDPs) are a class of highly flexible proteins whose characterization by NMR spectroscopy is complicated by severe spectral overlaps. The development of experiments designed to facilitate the sequence-specific assignment procedure is thus very important to improve the tools for the characterization of IDPs and thus to be able to focus on IDPs of increasing size and complexity. Here, we present and describe the implementation of a set of novel ¹H-detected 5D experiments, (HACA)CON(CACO)NCO(CA)HA, BT-(H)NCO(CAN)CONNH and BT-HN(COCAN)CONNH, optimized for the study of highly flexible IDPs that exploit the best resolved correlations, those involving the carbonyl and nitrogen nuclei of neighboring amino acids, to achieve sequence-specific resonance assignment. Together with the analogous recently proposed pulse schemes based on ¹³C detection, they form a complete set of experiments for sequence-specific assignment of highly flexible IDPs. Depending on the particular sample conditions (concentration, lifetime, pH, temperature, etc.), these experiments present certain advantages and disadvantages that will be discussed. Needless to say, that the availability of a variety of complementary experiments will be important for accurate determination of resonance frequencies in complex IDPs.
authors.affiliation: CERM and Department of Chemistry, University of Florence, Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Florence, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Piai
authors.forename: Tomáš
authors.initials: T
authors.lastname: Hošek
authors.forename: Leonardo
authors.initials: L
authors.lastname: Gonnelli
authors.forename: Anna
authors.initials: A
authors.lastname: Zawadzka-Kazimierczuk
authors.forename: Wiktor
authors.initials: W
authors.lastname: Koźmiński
authors.forename: Bernhard
authors.initials: B
authors.lastname: Brutscher
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Bermel
authors.forename: Roberta
authors.initials: R
authors.lastname: Pierattelli
authors.forename: Isabella C
authors.initials: IC
authors.lastname: Felli
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D064267:Intrinsically Disordered Proteins / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D013696:Temperature
chemical_list: D002247:Carbon Isotopes; D064267:Intrinsically Disordered Proteins