Renaturation of bovine erythrocyte carbonic anhydrase B denatured by acid, heat, and detergent

McCoy, Leslie F.; Wong, K.P.

doi:10.1021/bi00514a012

Cited by 50 publications

(58 citation statements)

References 33 publications

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“…Fig.1 reveals that the native structure of HCAB has about the same structural features as its bovine analog [20]. The far-UV CD spectrum is compatible with little helical order and about 35Yi p- structure while the characteristic near-UV CD bands originate from the aromatic amino acid clustering in the tertiary structure.…”

Section: Acid-induced Conformation Of Hcabmentioning

confidence: 88%

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The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

Jagannadham

Balasubramanian

1985

FEBS Letters

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The acid-induced and guanidinium chloride-induced conformational transitions in human carbonic anhydrase B have beerranalyzed. A structural form was detected at pH 3, which has a higher secondary structural order than the native enzyme but little tertiary structure. The enzyme dissolved in an intermediate concentration of the denaturant guanidinium chloride (1 M at pH 7.5) also adopts a similar conformational state. This form, denoted as the intermediate form I, possesses most of the characteristics defined for the molten globular state of globular proteins and might serve as the embryonic structural intermediate during the selforganization of the protein into its functional native form. Globular protein renaturation Molten globular form

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Section: Acid-induced Conformation Of Hcabmentioning

confidence: 88%

“…The enzyme aggregates and precipitates when one attempts to renature it from the acid-expanded form directly, just as the bovine analog does [20]. …”

Section: Time Dependence Of Structural Transitionsmentioning

confidence: 99%

The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

Jagannadham

Balasubramanian

1985

FEBS Letters

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“…It aggregates during renaturation when denatured with heat 26 or acid, 27 or when incubated in intermediate concentrations (1-3 M) of guanidinium chloride (GuHCl). [28][29][30] For example, Hammarström et al observed low (<10%) recovery of activity of human carbonic anhydrase (HCAII) when it was renatured by dilution from 2 to 0.3 M GuHCl, but high (>80%) recovery of activity when diluted from 6 M GuHCl to 0.3 M. 30 McCoy et al 26 also observed aggregation of BCA in refolding of the protein, denatured in a solution of 0.1% SDS and renatured with dialysis. 31 We believe that irreversible aggregation interferes with refolding of BCA (and BCA-Ac 18 ) in intermediate concentrations of SDS and prevents equilibration (eq 1).…”

Section: Why Do We Not Achieve Equilibration Between Native and Denatmentioning

confidence: 99%

Peracetylated Bovine Carbonic Anhydrase (BCA-Ac₁₈) Is Kinetically More Stable than Native BCA to Sodium Dodecyl Sulfate

2006

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Bovine carbonic anhydrase (BCA) and its derivative with all lysine groups acetylated (BCA-Ac 18 ) have different stabilities toward denaturation by sodium dodecyl sulfate (SDS). This difference is kinetic: BCA-Ac 18 denatures more slowly than BCA by several orders of magnitude over concentrations of SDS ranging from 2.5 to 10 mM. The rates of renaturation of BCA-Ac 18 are greater than those of BCA, when these proteins are allowed to refold from a denatured state ([SDS] ) 10 mM) to a folded state ([SDS] ) 0.1 to 1.5 mM). On renaturation, the yields of the correctly folded protein (either BCA or BCA-Ac 18 ) decrease with increasing concentration of SDS. At intermediate concentrations of SDS (from 0.7 to 2 mM for BCA, and from 1.5 to 2 mM for BCA-Ac 18 ), both unfolding and refolding of the proteins are too slow to be observed; an alternative processs probably aggregationscompetes with refolding of the denatured proteins at those intermediate concentrations.Because it is experimentally impractical to prove equilibrium, it is not possible to establish whether there is a difference in the thermodynamics of unfolding/refolding between BCA and BCA-Ac 18 .

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“…CD studies by manual mixing have shown [6,7] that the secondary structure of carbonic anhydrase is restored in less than 100s. CD measurements with the stopped-flow technique have not yet been performed.…”

Section: Febs Letters November 1987mentioning

confidence: 99%

“…In particular, it has been shown that refolding of carbonic anhydrase B proceeds through an intermediate state [4][5][6][7] which has been identified as a 'molten globule' state [7]. However, the refolding kinetics was studied at time intervals >100 s, therefore the formation of early kinetic intermediates could not be followed.…”

Section: Introductionmentioning

confidence: 99%

Sequential mechanism of refolding of carbonic anhydrase B

et al. 1987

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The kinetics of refolding of bovine carbonic anhydrase B was studied by a variety of methods over a wide range of times (from milliseconds to hours). It has been shown that protein refolding proceeds through three stages. At the first stage (tl/2~0.03 s) hydrophobic clusters and a compact state of the chain are formed. At the second stage (tl,'2 ~ 140 s) hydrophobic clusters are desolvated and the rigid native-like hydrophobic core is formed. At the third stage (tl/2 ~ 600 s) the native active protein is formed.

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Renaturation of bovine erythrocyte carbonic anhydrase B denatured by acid, heat, and detergent

Cited by 50 publications

References 33 publications

The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

Peracetylated Bovine Carbonic Anhydrase (BCA-Ac₁₈) Is Kinetically More Stable than Native BCA to Sodium Dodecyl Sulfate

Sequential mechanism of refolding of carbonic anhydrase B

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Renaturation of bovine erythrocyte carbonic anhydrase B denatured by acid, heat, and detergent

Cited by 50 publications

References 33 publications

The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

The molten globular intermediate form in the folding pathway of human carbonic anhydrase B

Peracetylated Bovine Carbonic Anhydrase (BCA-Ac18) Is Kinetically More Stable than Native BCA to Sodium Dodecyl Sulfate

Sequential mechanism of refolding of carbonic anhydrase B

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Peracetylated Bovine Carbonic Anhydrase (BCA-Ac₁₈) Is Kinetically More Stable than Native BCA to Sodium Dodecyl Sulfate