title: Acquisition of native beta-strand topology during the rapid collapse phase of protein folding.
abstract: The 98 residue C-terminal domain of the cell-surface receptor protein CD2 (CD2.D1) has a beta-sandwich fold belonging to the immunoglobulin superfamily but lacking the usual disulfide bridges. Kinetic studies on the folding/unfolding of CD2.D1 reveal that folding proceeds through a rapidly formed intermediate state [Parker, M. J., & Clarke, A. R. (1997) Biochemistry 36, 5786-5794]. To characterize the structural properties of this intermediate we have performed a series of amide hydrogen exchange studies using the pH competition method, in which folding and exchange are initiated simultaneously. The complex beta-sheet topology of this molecule makes it an ideal object for examining the acquisition of backbone hydrogen bonds made between sequence-local and sequence-distant segments of the chain during folding. The pattern of protected amides in the intermediate reveal that the essential features of the beta-sheet topology of CD2.D1 are defined early in the folding pathway, before the development of intimate side chain interactions characteristic of the native state. The results are discussed in light of current issues concerning the mechanistic relevance of kinetic protein folding intermediates.
authors.affiliation: Department of Biochemistry, University of Bristol, School of Medical Sciences, University Walk, Bristol BS8 1TD, U.K.
authors.forename: M J
authors.initials: MJ
authors.lastname: Parker
authors.forename: C E
authors.initials: CE
authors.lastname: Dempsey
authors.forename: M
authors.initials: M
authors.lastname: Lorch
authors.forename: A R
authors.initials: AR
authors.lastname: Clarke
mesh_terms: D000577:Amides; D000818:Animals; D018801:CD2 Antigens / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D051381:Rats; D012995:Solubility; D013816:Thermodynamics
chemical_list: D000577:Amides; D018801:CD2 Antigens