 Transition state structures in two-state folding are not experimentally accessible (by definition they are the least populated along the reaction coordinate), but the folding sub-ensembles in downhill folding processes are theoretically distinguishable by spectroscopy.The 40-residue protein BBL, which is an independently folding domain from the E2 subunit of the 2-oxoglutarate dehydrogenase multi-enzyme complex of E. coli, has been experimentally shown to fold globally downhill.Also, a mutant of lambda repressor protein has been shown to shift from downhill to two-state upon changing the temperature/solvent conditions.