Three-phase partitioning (TPP) is carried out by mixing ammonium sulfate and t-butanol to obtain organic phase, interfacial precipitate and aqueous phase. It is shown that TPP of an 8 M urea/100 mM dithiothreitol-denatured xylanase preparation resulted in simultaneous renaturation and purification. This integrated novel approach gave recovery of 93% enzyme activity with 21-fold purification. Th...

We have studied nucleic acid double helix destabilization mediated by purified calf helix-unwinding proteins, measuring ultraviolet hyperchromicity to detect helix melting. Both calf unwinding protein 1 (UP1) and a high salt eluting protein fraction are found to depress strongly the helix melting temperature (Tm) of the synthetic alternating copolymers poly[d(AT)] and poly[r(AU)], indicating th...

Using a double antibody sandwich ELISA we examined the heavy chain isoform composition of myosin molecules isolated from chicken pectoralis major muscle during different stages of development. At 2- and 40-d posthatch, when multiple myosin heavy chain isoforms are being synthesized, we detected no heterodimeric myosins, suggesting that myosins are homodimers of the heavy chain subunit. Chymotry...

Structural characteristics of numerous globular proteins in the denatured state have been reviewed using literature data. Recent more precise experiments show that in contrast to the conventional standpoint, proteins under strongly denaturing conditions do not unfold completely and adopt a random coil state, but contain significant residual ordered structure. These results cast doubt on the bas...

The possibility of engineering near-infrared fluorescent proteins and biosensors from bacterial phytochrome photoreceptors (BphPs) has led to substantial interest in this family of proteins. The near-infrared fluorescent proteins have allowed non-invasive bio-imaging of deep tissues and whole organs in living animals. BphPs and derived near-infrared fluorescent proteins contain a structural ele...