title: Two immunoglobulin tandem proteins with a linking β-strand reveal unexpected differences in cooperativity and folding pathways.
abstract: The study of the folding of single domains, in the context of their multidomain environment, is important because more than 70% of eukaryotic proteins are composed of multiple domains. The structures of the tandem immunoglobulin (Ig) domain pairs A164-A165 and A168-A169, from the A-band of the giant muscle protein titin, reveal that they form tightly associated domain arrangements, connected by a continuous β-strand. We investigate the thermodynamic and kinetic properties of these tandem domain pairs. While A164-A165 apparently behaves as a single cooperative unit at equilibrium, unfolding without the accumulation of a large population of intermediates, domains in A168-A169 behave independently. Although A169 appears to be stabilized in the tandem protein, we show that this is due to nonspecific stabilization by extension. We elucidate the folding and unfolding pathways of both tandem pairs and show that cooperativity in A164-A165 is a manifestation of the relative refolding and unfolding rate constants of each individual domain. We infer that the differences between the two tandem pairs result from a different pattern of interactions at the domain/domain interface.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
authors.forename: Annette
authors.initials: A
authors.lastname: Steward
authors.forename: Qing
authors.initials: Q
authors.lastname: Chen
authors.forename: Robert I
authors.initials: RI
authors.lastname: Chapman
authors.forename: Madeleine B
authors.initials: MB
authors.lastname: Borgia
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Rogers
authors.forename: Alexsandra
authors.initials: A
authors.lastname: Wojtala
authors.forename: Matthias
authors.initials: M
authors.lastname: Wilmanns
authors.forename: Jane
authors.initials: J
authors.lastname: Clarke
mesh_terms: D064211:Connectin; D004926:Escherichia coli / Q000378:metabolism; D007136:Immunoglobulins / Q000737:chemistry*; D007700:Kinetics; D008958:Models, Molecular; D009124:Muscle Proteins / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D011494:Protein Kinases / Q000737:chemistry; D017434:Protein Structure, Tertiary; D013816:Thermodynamics
chemical_list: D064211:Connectin; D007136:Immunoglobulins; D009124:Muscle Proteins; D011494:Protein Kinases