Non-random-coil behavior as a consequence of extensive PPII structure in the denatured state.
Unfolded proteins may contain a native or nonnative residual structure, which has important implications for the thermodynamics and kinetics of folding, as well as for misfolding and aggregation diseases. However, it has been universally accepted that residual structure should not affect the global size scaling of the denatured chain, which obeys the statistics of random coil polymers. Here we use a single-molecule optical technique--fluorescence correlation spectroscopy--to probe the denatured state of a set of repeat proteins containing an increasing number of identical domains, from 2 to 20. The availability of this set allows us to obtain the scaling law for the unfolded state of these proteins, which turns out to be unusually compact, strongly deviating from random coil statistics. The origin of this unexpected behavior is traced to the presence of an extensive nonnative polyproline II helical structure, which we localize to specific segments of the polypeptide chain. We show that the experimentally observed effects of polyproline II on the size scaling of the denatured state can be well-described by simple polymer models. Our findings suggest a hitherto unforeseen potential of nonnative structure to induce significant compaction of denatured proteins, significantly affecting folding pathways and kinetics.
Cortajarena
Aitziber L
AL
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA.
Lois
Gregg
G
Sherman
Eilon
E
O'Hern
Corey S
CS
Regan
Lynne
L
Haran
Gilad
G
D002942:Circular Dichroism; D003198:Computer Simulation; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry; D020449:Repetitive Sequences, Amino Acid; D012235:Rhodamines; D013057:Spectrum Analysis; D014783:Viscosity
D010455:Peptides; D011506:Proteins; D012235:Rhodamines; C026188:rhodamine 6G; C011083:polyproline