Intrinsic fluorescence and activity of the original HsTIM, P1 and P2 without and with 3 M urea.

The λmax of the intrinsic fluorescence and specific activity (µmol/min/mg) of the purified native HsTIM in the absence and presence of 3 M urea are shown in the first two lines.aData of P1 and P2 eluted from SEC in urea buffer and maintained the urea buffer.bData of P1 and P2 after urea was removed by extensive dialysis in 100 mM triethanolamine, 10 mM EDTA and 1 mM DTT (pH7.4).cData with the latter enzymes after 3 M urea was added back again. Activities were measured with 1 mM glyceraldehyde 3-phosphate.The activity of P1 in the presence of 3 M urea could not be accurately measured because it is denatured in buffer with 3 M urea. After it is transferred to the assay media (which has no urea), it undergoes progressive reactivation.