Complete amino acid sequence of rabbit muscle glycogen phosphorylase.

Titani, Koiti; Koide, Atsushi; Hermann, Jacques; Ericsson, Lowell H.; Kumar, Santosh; Wade, Roger D.; Walsh, Kenneth A.; Neurath, Hans; Fischer, Edmond H.

doi:10.1073/pnas.74.11.4762

Cited by 221 publications

(80 citation statements)

References 27 publications

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“…In order to calculate the degree of phosphorylation, the molecular weights of phosphorylase and synthase subunits were taken as 98000 [26] and 88000 [21] respectively. Protein concentrations were measured spectrophotometrically using absorbance indices, A;$ nm, of 13.1 (phosphorylase) [27] and 13.4(glycogen synthase) [21].…”

Section: Phosphorylation Of Glycogen Synthase a And Phosphorylase B Bmentioning

confidence: 99%

Glycogen Synthase Kinase-2 and Phosphorylase Kinase Are the Same Enzyme

Embi¹,

Rylatt²,

Cohen³

1979

Eur J Biochem

112

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Homogeneous preparations of phosphorylase kinase from rabbit skeletal muscle catalyse a calcium-dependent phosphorylation of glycogen synthase a isolated from the same tissue. The calcium-dependent glycogen synthase kinase activity copurifies with phosphorylase kinase throughout the standard procedure for the isolation of the latter enzyme. At the final step of the purification, gel filtration on Sepharose 4B, the elution profiles for glycogen synthase kinase and phosphorylase kinase activities are identical. ICR/IAn mice, which lack muscle phosphorylase kinase activity, do not contain detectable calcium-dependent glycogen synthase kinase activity. These results indicate that the calcium-dependent phosphorylation of glycogen synthase is catalysed by phosphorylase kinase and not by another calcium-dependent protein kinase that might be contaminating the preparation.The phosphorylation of glycogen synthase a by phosphorylase kinase reaches a plateau in the range 0.73 0.1 molecules of phosphate incorporated per subunit and is accompanied by a 2-fold decrease in the activity in the absence of glucose 6-phosphate. The phosphorylation takes place on a unique serine residue located seven amino acids from the N-terminus of the polypeptide chain. The amino acid sequence surrounding serine-7 is similar to the amino acid sequence surrounding the phosphoserine in phosphorylase a.Glycogen synthase kinase-2 is the name given to a protein kinase present as a trace contaminant in highly purified preparations of glycogen synthase [Nimmo, H. G. and Cohen, P. (1974) FEBS Lett. 47, 162 -1671. Glycogen synthase kinase-2 also phosphorylates serine-7 exclusively, and evidence is presented which demonstrates that glycogen synthase kinase-2 is merely a modified form of phosphorylase kinase which has lost its ability to be regulated by calcium ions at pH 6.8.The rate of phosphorylation of glycogen synthase a by phosphorylase kinase is 2 -3-fold slower than the rate of phosphorylation of phosphorylase h when identical concentrations of the two protein substrates are used (6 pM). At physiological concentrations of glycogen synthase (0.3 mg/ml) and phosphorylase (8.0 mgiml), the time required for half-maximal phosphorylation of each enzyme by phosphorylase kinase is similar. These results suggest that the phosphorylation of glycogen synthase by phosphorylase kinase may be physiologically significant, and the implications of these findings are considered.

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Section: Phosphorylation Of Glycogen Synthase a And Phosphorylase B Bmentioning

confidence: 99%

Glycogen Synthase Kinase-2 and Phosphorylase Kinase Are the Same Enzyme

Embi¹,

Rylatt²,

Cohen³

1979

Eur J Biochem

112

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“…The numbering scheme is the same as for the protein sequence for residues 1 (Ser) to 307 (Be). After Be 307, the insertion of an isoleucine residue causes the subsequent amino acid sequence shown here to be incremented by one over the previously published protein sequence [3]. Such discrepancies are occasionally found between primary structures determined from protein and DNA sequencing.…”

Section: Resultsmentioning

confidence: 80%

“…We have compared the phosphorylase amino acid sequence deduced from cDNA sequencing with that determined by protein sequencing [3]. An additional amino acid, isoleucine, has been introduced at position 308 in the amino acid sequence deduced from cDNA.…”

Section: Resultsmentioning

confidence: 99%

“…X-ray crystallographic analysis [2] and the amino acid sequence [3] of this enzyme have led to a three-dimensional model of the protein structure and proposals for catalytic and regulatory m~hanisms. A powerful method for testing such proposals and thus advancing our understanding of the molecular mechanisms of phosphorylase function is to analyze the structure and enzymatic properties of appropriately mutated enzyme.…”

mentioning

confidence: 99%

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Complete cDNA sequence for rabbit muscle glycogen phosphorylase

Nakano¹,

Hwang²,

Fletterick³

1986

FEBS Letters

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The cDNA for the nearly full-length rabbit muscle giycogen phospho~lase mRNA has been isolated and sequenced. The cDNA is rich in G and C nucleotides. This feature is especially striking at the 3rd position of codons, where 86% of the 843 amino acid codons terminate with G or C. Methionine, presumably the initiation residue, is found at position-1, suggesting that the removal of only a single methionine residue precedes the amino-terminal acetylation at serine. Eight differences between the deduced amino acid sequence and the previously determined protein sequence are discussed. Amino acid sequence

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“…Human muscle extracts showed a single component with Mr of -110 000. In view of the results obtained in sedimentation experiments and the known M, of rabbit muscle phosphorylase b (dimer, 194 664) [6], it seems likely that some of the M,-values obtained in this way are low, possibly because of interactions of the phosphorylases with the gel matrix of Sephacryl. The very low apparent M,-values for Ph II and the rabbit brain enzyme point to significant dissociation to monomer at these low concentrations (-1 EU/ml extract):…”

Section: Methodsmentioning

confidence: 92%