Comparative properties of glycogen phosphorylase. VIII. Phosphorylase from dogfish skeletal muscle. Purification and a comparison of its physical properties to those of rabbit muscle phosphorylase

Cohen, Philip; Duewer, Theresa; Fischer, Edmond H.

doi:10.1021/bi00790a005

Cited by 149 publications

(85 citation statements)

References 56 publications

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“…Glucose is known to promote dissociation of the tetrameric rabbit muscle phosphorylase a to dimers (Wang et al, 1965;Helmreich et al, 1967;Withers et al, 1979). The values of 8.4-9.4 S and 13.0-14.2 S obtained in this study closely correspond to those of a dimer and a tetramer, respectively (Wang et al, 1965;Cohen et al, 1971;Withers et al, 1979Withers et al, , 1982b. The following results were obtained.…”

Section: Kineticssupporting

confidence: 74%

N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

et al. 1995

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Structure-based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to a-D-glucose (Ki = 1.7 mM). The best inhibitor in the class of N-acyl derivatives of P-D-glucopyranosylamine, N-acetyl-P-D-glucopyranosylamine (I-GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1-GlcNAc acts as a competitive inhibitor for both the b (K, = 32 pM) and the a ( K , = 35 PM) forms of the enzyme with respect to glucose I-phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1-GlcNAc was able to induce dissociation of tetrameric phosphorylase a and stabilization of the dimeric T-state conformation. Co-crystals of the phosphorylase b-I-GlcNAc-IMP complex were grown in space group P432,2, with native-like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1 %, for data between 8 and 2.3 A resolution. I-GlcNAc binds tightly at the catalytic site of T-state phosphorylase b at approximately the same position as that of a-D-glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T-state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T-state phosphorylase b-I-GlcNAc-IMP complex structure is overall similar to the T-state phosphorylase b-a-D-glucose complex structure. The structure of the I-GlcNAc complex provides a rational for the biochemical properties of the inhibitor.

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Section: Kineticssupporting

confidence: 74%

N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

et al. 1995

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“…Table 1 shows the sedimentation constants ( s~~,~) determined for GPb (4 mg/mL) in the presence of ammonium sulfate and various ligands. The values of 7.9-8.8s and 12.4-14.1s obtained in this study closely correspond to those of a dimer and tetramer, respectively (Wang et al, 196513;Cohen et al, 1971;Withers et al, 1979Withers et al, , 1982. The following results were obtained: (1) In the absence of AMP, GPb sedimented 35% as a dimer (s20,w = 8.7s) and 65% as a tetramer (s20,w = 14.0s).…”

Section: Ultracentrifugationsupporting

confidence: 85%

Kinetic properties of tetrameric glycogen phosphorylase b in solution and in the crystalline state

et al. 1992

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“…Protein concentration of immunoglobulin fraction or of purified antibodies was determined either by the method of Lowry et al [16] or by absorption at 280 nm using rabbit immunoglobulin as standard. The molecular weight of the dimeric enzyme was taken as 200000 [17] while the average molecular weights for antibodies was assumed equal to 150000. Absorption spectra were taken by means of a PerkinElmer 356 Double Beam UV-Visible recording spectrophotometer.…”

Section: Methodsmentioning

confidence: 99%

Antibodies and Autoantibodies of Glycogen Phosphorylase b: Inactivation of Pig and Rabbit Enzymes

Tzartos

Evangelopoulos

1977

European Journal of Biochemistry

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Pig skeletal muscle glycogen phosphorylase b was purified using ammonium sulfate fractionation, DEAE-Sephadex A-50 and Sephadex G-200 column chromatography. The purified enzyme was used to immunize rabbits in the presence or in the absence of complete Freund adjuvant.Antibodies against pig phosphorylase in pure form were isolated from rabbit antisera using insoluble immunoadsorbents of pig phosphorylase. Autoantibodies against the rabbit enzyme were obtained from the same antisera using insoluble immunoadsorbents of rabbit phosphorylase. Complete inactivation of pig phosphorylase was accomplished by an antibody/enzynie molar ratio equal to 4 and autoantibody/enzyme molar ratio equal to 130. Complete inactivation of rabbit phosphorylase was accomplished by an antibody/enzyme molar ratio equal to 250 and autoantibody/enzyme molar ratio equal to 160. Passive haemagglutination technique gave positive results with minimum amounts of 0.02 pg/ml and 0.8 pg/ml for pig and rabbit phosphorylase respectively.Kinetic experiments have shown that antibodies and autoantibodies act as noncompetitive inhibitors of both enzymes with respect to AMP and glucose l-phosphate but exhibit a mixed type of inhibition with respect to glycogen. When glycogen hydrolysates were used as substrate in place of intact glycogen molecules a pronounced decrease in the inhibitory capacity of antienzyme on the enzyme was demonstrated.

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Comparative properties of glycogen phosphorylase. VIII. Phosphorylase from dogfish skeletal muscle. Purification and a comparison of its physical properties to those of rabbit muscle phosphorylase

Cited by 149 publications

References 56 publications

N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

N‐acetyl‐β‐D‐glucopyranosylamine: A potent T‐state inhibitor of glycogen phosphorylase. A comparison with α‐D‐glucose

Kinetic properties of tetrameric glycogen phosphorylase b in solution and in the crystalline state

Antibodies and Autoantibodies of Glycogen Phosphorylase b: Inactivation of Pig and Rabbit Enzymes

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