Kinetics of interaction of partially folded proteins with a hydrophobic dye: Evidence that molten globule character is maximal in early folding intermediates

Interaction with 8‐anilino‐1‐naphthalenesulfonate (ANS) is widely used to detect molten globule states of proteins. We have found that even with stable partially folded states, the development of the fluorescence enhancements resulting from such interactions can be relatively slow and kinetically complex. This is probably because initial binding of the dye can induce subsequent changes in the protein structure, so that the ultimate resulting fluorescence enhancement is not necessarily a good, nonperturbing probe of the preexisting state of the protein. When ANS is used to study folding mechanisms the problem is compounded by the difficulty of distinguishing effects due to the development of dye interactions from those due to the changing populations of folding intermediates.

[1]  R. Yada,et al.  Evidence for a molten globule state in an oligomeric plant protein , 1997 .

[2]  J. Beechem,et al.  Local and global dynamics during the folding of Escherichia coli dihydrofolate reductase by time-resolved fluorescence spectroscopy. , 1995, Biochemistry.

[3]  C. Matthews,et al.  Development of nonpolar surfaces in the folding of Escherichia coli dihydrofolate reductase detected by 1-anilinonaphthalene-8-sulfonate binding. , 1994, Biochemistry.

[4]  A. Fink,et al.  Protein conformational changes induced by 1,1'-bis(4-anilino-5-naphthalenesulfonic acid): preferential binding to the molten globule of DnaK. , 1994, Biochemistry.

[5]  C. Dobson,et al.  Tertiary interactions in the folding pathway of hen lysozyme: kinetic studies using fluorescent probes. , 1994, Biochemistry.

[6]  M. Cocco,et al.  The native state of apomyoglobin described by proton NMR spectroscopy: Interaction with the paramagnetic probe HyTEMPO and the fluorescent dye ANS , 1994, Protein science : a publication of the Protein Society.

[7]  J. King,et al.  Folding of the phage P22 coat protein in vitro. , 1993, Biochemistry.

[8]  P. Leadlay,et al.  Purification and separation of holo- and apo-forms of Saccharopolyspora erythraea acyl-carrier protein released from recombinant Escherichia coli by freezing and thawing. , 1993, The Biochemical journal.

[9]  Matthews Cr,et al.  Structure and stability of an early folding intermediate of Escherichia coli trp aporepressor measured by far-UV stopped-flow circular dichroism and 8-anilino-1-naphthalene sulfonate binding. , 1993 .

[10]  A. Chaffotte,et al.  Kinetic resolution of peptide bond and side chain far-UV circular dichroism during the folding of hen egg white lysozyme. , 1992, Biochemistry.

[11]  N. A. Rodionova,et al.  Study of the “molten globule” intermediate state in protein folding by a hydrophobic fluorescent probe , 1991, Biopolymers.

[12]  S Sugai,et al.  An early immunoreactive folding intermediate of the tryptophan synthase β2 subunit is a ‘molten globule’ , 1990, FEBS letters.

[13]  O. Ptitsyn,et al.  Evidence for a molten globule state as a general intermediate in protein folding , 1990, FEBS letters.

[14]  P. Horowitz,et al.  Reversible folding of rhodanese. Presence of intermediate(s) at equilibrium. , 1989, The Journal of biological chemistry.

[15]  C M Dobson,et al.  Characterization of a partly folded protein by NMR methods: studies on the molten globule state of guinea pig alpha-lactalbumin. , 1989, Biochemistry.

[16]  O. Ptitsyn,et al.  Sequential mechanism of refolding of carbonic anhydrase B , 1987, FEBS letters.

[17]  S Sugai,et al.  Rapid formation of secondary structure framework in protein folding studied by stopped‐flow circular dichroism , 1987, FEBS letters.

[18]  R. Clegg,et al.  Diffusion-controlled association of a dye, 1-anilinonaphthalene-8-sulfonic acid, to a protein, bovine serum albumin, using a fast-flow microsecond mixer and stopped-flow. , 1987, Biophysical chemistry.

[19]  A. Labhardt.  Kinetic circular dichroism shows that the S-peptide alpha-helix of ribonuclease S unfolds fast and refolds slowly. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M. Kronman,et al.  Binding of naphthalene dyes to the N and A conformers of bovine α-lactalbumin , 1982 .

[21]  H. Nakatani,et al.  Kinetic studies on binding of bovine serum albumin with 1‐anilino‐8‐naphthalene sulfonate , 1974, FEBS letters.

[22]  L. Stryer,et al.  The interaction of a naphthalene dye with apomyoglobin and apohemoglobin. A fluorescent probe of non-polar binding sites. , 1965, Journal of molecular biology.

[23]  S. Radford,et al.  Probing the structure of folding intermediates , 1994 .

[24]  L. Serrano.  Protein engineering and the dissection of protein folding pathways , 1994 .

[25]  C. Matthews,et al.  Structure and stability of an early folding intermediate of Escherichia coli trp aporepressor measured by far-UV stopped-flow circular dichroism and 8-anilino-1-naphthalene sulfonate binding. , 1993, Biochemistry.

[26]  C. Matthews,et al.  Pathways of protein folding. , 1993, Annual review of biochemistry.

[27]  P. S. Kim,et al.  Intermediates in the folding reactions of small proteins. , 1990, Annual review of biochemistry.