Interaction of muscle glycogen phosphorylase with pyridoxal 5′-methylenephosphonate

Vidgoff, J.M.; Pocker, Anna; Hullar, T. L.; Fischer, Edmond H.

doi:10.1016/0006-291x(74)90819-5

Cited by 24 publications

(19 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Chemical modification studies have directed attention to the phosphate group of pyridoxal-P (4,5,15,24). This phosphate is not transferred during catalysis (1), but its role as a proton donor/acceptor is suggested by the fact that maximal enzyme activity occurs at a pH very near the pKa of pyridoxal-P for native phosphorylase and near the pKa of pyridoxal 5'-methylenephosphonate for the corresponding phosphorylase derivative (24).…”

Section: Discussionmentioning

confidence: 99%

31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate.

Feldmann¹,

Hull²

1977

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

ABSTRACT31P nuclear magnetic resonance (NMR) at 72.8 MHz has been used to study glycogen phosphorylase from rabbit muscle (1,4-a-D-glucan:orthophosphate a-glucosyltransferase, EC 2.4.1.1) at concentrations as low as 25 mg/ml, using a WH-180 wide-bore superconducting spectrometer. The use of a thio analogue for 5'-AMP and arsenate for inorganic phosphate allowed the observation of three distinct forms of enzyme-bound pyridoxal 5'-phosphate at -0.2 ppm (Form I), -2 to -3 ppm (Form II, and -3.5 ppm (Form III) relative to triethylphosphate. Conversion of I to III occurs by activation of phosphorylase either by formation-of a ternary complex of phosphorylase b with effector and arsenate or, more efficiently, by direct phosphorylation to give the a form of the enzyme. The ionization state and exposure to solvent of each of the three forms is inferred from the 31P NMR data.Pyridoxal 5'-phosphate is an essential constituent of all known a-glucan phosphorylases (1,4-a-D-glucan:orthophosphate aglucosyltransferase, EC 2.4.1.1). Reconstitution experiments with pyridoxal-P analogues (1-3) indicate the importance of the phosphate moiety in pyridoxal-P function but do not provide information on its precise role (4, 5). 31P nuclear magnetic resonance (NMR) spectroscopy gives specific information on the ionization state and the pKa values of pyridoxal-P bound to the enzyme. In addition, one can observe all other phosphorus-containing ligands involved in phosphorylase catalysis, as well as the phosphoserine group that regulates the activity of phosphorylase by interconversion of the b and a forms (6).However, the 31P NMR signals for Pi, glucose 1-phosphate, AMP, phosphoserine, and pyridoxal-P all lie in a very narrow range of ca 4 ppm. Therefore, we have substituted adenosine 5'-O-thiomonophosphate (AMP-S) for AMP and arsenate for phosphate. Adenosine 5'-O-(3-thio)triphosphate (ATP-y-S) was used to prepare phosphorylase a containing thiophosphoserine residues. Thus, it was possible to separate the signals of the various ligands and to clearly observe the pyridoxal-P resonance, which monitors the protein conformational change induced by the effector AMP or by the phosphoserine group.

show abstract

Section: Discussionmentioning

confidence: 99%

31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate.

Feldmann¹,

Hull²

1977

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…ties. The results of reconstitution studies using various PLP analogues have demonstrated the importance of the S'-phosphate group of PLP in phosphorylase action (Shaltiel et al, 1969b;Pfeuffer et al, 1972a,b;Vidgoff et al, 1974;Parrish et al, 1977). The results of recent X-ray crystallographic studies on rabbit muscle phosphorylase a have shown that PLP is buried inside the protomer and its S'-phosphate group is located adjacent to the substrate site (Sygusch et al, 1977).…”

mentioning

confidence: 99%

Characterization of the pyridoxal phosphate site in glycogen phosphorylase b from rabbit muscle

Shimomura¹,

Fukui²

1978

Biochemistry

View full text Add to dashboard Cite

show abstract

“…With the notable exception of S'-deoxypyridoxalmethylenephosphonic acid (Fischer et al, 1970;Vidgoff et al, 1974) and S'-deoxypyridoxalphosphonic acid (Graves and Wang, 1972), all other modifications of the cofactor at the 5' position have resulted in proteins reported to be catalytically inactive. Thus, it was concluded that the 5' position was critical for activity, and on the basis of studies with pyridoxal S'-phosphate monomethyl ester reconstituted phosphorylase, a proton transfer 4824 BIOCHEMISTRY.…”

mentioning

confidence: 99%

Effect of phosphate analogs on the activity of pyridoxal reconstituted glycogen phosphorylase

Parrish¹,

Uhing²,

Graves³

1977

Biochemistry

View full text Add to dashboard Cite

Interaction of muscle glycogen phosphorylase with pyridoxal 5′-methylenephosphonate

Cited by 24 publications

References 15 publications

31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate.

31P nuclear magnetic resonance studies of glycogen phosphorylase from rabbit skeletal muscle: ionization states of pyridoxal 5'-phosphate.

Characterization of the pyridoxal phosphate site in glycogen phosphorylase b from rabbit muscle

Effect of phosphate analogs on the activity of pyridoxal reconstituted glycogen phosphorylase

Contact Info

Product

Resources

About