The denaturation of rabbit muscle phosphorylase <i>b</i> by guanidinium chloride

Price, Nicholas C.; Stevens, Evelyn

doi:10.1042/bj2130595

Cited by 11 publications

(4 citation statements)

References 24 publications

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“…Common examples are the following: (1) formation of inclusion bodies in E. coli that have been engineered by recombinant DNA technology to overexpress heterologous proteins (Uren, 1984); (2) precipitation of heat-treated globular proteins (Privalov, 1979); and (3) irreversibility of protein renaturation due to aggregation (Light, 1985). Apparent irreversibility of in vitro refolding was reported for tryptophanase (London et al, 1974), 0-galactosidase (Goldberg, 1972, catalytic subunit of aspartate transcarbamylase (Ghelis & Hervé, 1978), elastase (Zilber, 1979), the dehydrogenases (Jaenicke & Rudolph, 1983), rhodanese (Horowitz & Criscimagna, 1986), phosphorylase b (Price & Stevens, 1983), and antithrombin (Fish et al, 1985). For these proteins, the irreversibility of refolding was shown to occur only at critical concentrations of denaturant.…”

Section: Bremsmentioning

confidence: 97%

Solubility of different folding conformers of bovine growth hormone

Brems¹

1988

Biochemistry

120

116

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Section: Bremsmentioning

confidence: 97%

Solubility of different folding conformers of bovine growth hormone

Brems¹

1988

Biochemistry

120

116

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“…Under these conditions only very small changes in the secondary and tertiary structure of the enzyme had occurred as indicated by the far and near UV CD respectively [36] (see Fig. 5).…”

Section: Pyridoxal-5′-phosphatementioning

confidence: 99%

The Use of Circular Dichroism in the Investigation of Protein Structure and Function

Kelly

Price²

2000

CPPS

977

707

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Circular Dichroism (CD) relies on the differential absorption of left and right circularly polarised radiation by chromophores which either possess intrinsic chirality or are placed in chiral environments. Proteins possess a number of chromophores which can give rise to CD signals. In the far UV region (240-180 nm), which corresponds to peptide bond absorption, the CD spectrum can be analysed to give the content of regular secondary structural features such as α-helix and β-sheet. The CD spectrum in the near UV region (320-260 nm) reflects the environments of the aromatic amino acid side chains and thus gives information about the tertiary structure of the protein. Other non-protein chromophores such as flavin and haem moieties can give rise to CD signals which depend on the precise environment of the chromophore concerned. Because of its relatively modest resource demands, CD has been used extensively to give useful information about protein structure, the extent and rate of structural changes and ligand binding. In the protein design field, CD is used to assess the structure and stability of the designed protein fragments. Studies of protein folding make extensive use of CD to examine the folding pathway; the technique has been especially important in characterising "molten globule" intermediates which may be involved in the folding process. CD is an extremely useful technique for assessing the structural integrity of membrane proteins during extraction and characterisation procedures. The interactions between chromophores can give rise to characteristic CD signals. This is well illustrated by the case of the light harvesting complex from photosynthetic bacteria, where the CD spectra can be analysed to indicate the extent of orbital overlap between the rings of bacteriochlorophyll molecules. It is therefore evident that CD is a versatile technique in structural biology, with an increasingly wide range of applications.

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“…By studying effects of guanidinium hydrochloride on enzyme activity and structural integrity of rabbit muscle glycogen phosphorylase, Price and Stevens [19] concluded that this phosphorylase is much more sensitive to chemical denaturation than a number of other oligomeric proteins. A marked conformational flexibility of glycogen phosphorylase which seems to be required to achieve the complex regulatory properties of this enzyme [13][14][15][16][17][18], has been hypothesized to bring about the relatively low stability of the phosphorylase [19]. Large differences have recently been observed among individual α-glucan phosphorylases from bacteria, plant and mammals regarding the thermostability of enzyme activity [20].…”

Section: Introductionmentioning

confidence: 99%

Mechanism of thermal denaturation of maltodextrin phosphorylase from Escherichia coli

Griessler

D’Auria

Schinzel

et al. 2000

Biochemical Journal

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Maltodextrin phosphorylase from Escherichia coli (MalP) is a dimeric protein in which each $ 90-kDa subunit contains activesite pyridoxal 5h-phosphate. To unravel factors contributing to the stability of MalP, thermal denaturations of wild-type MalP and a thermostable active-site mutant (Asn-133 Ala) were compared by monitoring enzyme activity, cofactor dissociation, secondary structure content and aggregation. Small structural transitions of MalP are shown by Fourier-transform infrared spectroscopy to take place at $ 45 mC. They are manifested by slight increases in unordered structure and "H\#H exchange, and reflect reversible inactivation of MalP. Aggregation of the MalP dimer is triggered by these conformational changes and starts at $ 45 mC without prior release into solution of pyridoxal 5h-phosphate. It is driven by electrostatic rather than hydrophobic interactions between MalP dimers, and leads to irreversible inactivation of the enzyme. Aggregation is inhibited efficiently and specifically by oxyanions such as phosphate, and AMP

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The denaturation of rabbit muscle phosphorylase b by guanidinium chloride

Cited by 11 publications

References 24 publications

Solubility of different folding conformers of bovine growth hormone

Solubility of different folding conformers of bovine growth hormone

The Use of Circular Dichroism in the Investigation of Protein Structure and Function

Mechanism of thermal denaturation of maltodextrin phosphorylase from Escherichia coli

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