Glycoconjugate Journal

, Volume 34, Issue 4, pp 563–574 | Cite as

Probing the catalytic site of rabbit muscle glycogen phosphorylase using a series of specifically modified maltohexaose derivatives

  • Makoto Nakamura
  • Yasushi MakinoEmail author
  • Chika Takagi
  • Tohru Yamagaki
  • Masaaki Sato
Original Article


Glycogen phosphorylase (GP) is an allosteric enzyme whose catalytic site comprises six subsites (SG1, SG−1, SG−2, SG−3, SG−4, and SP) that are complementary to tandem five glucose residues and one inorganic phosphate molecule, respectively. In the catalysis of GP, the nonreducing-end glucose (Glc) of the maltooligosaccharide substrate binds to SG1 and is then phosphorolyzed to yield glucose 1-phosphate. In this study, we probed the catalytic site of rabbit muscle GP using pyridylaminated-maltohexaose (Glcα1–4Glcα1–4Glcα1–4Glcα1–4Glcα1–4GlcPA, where GlcPA = 1-deoxy-1-[(2-pyridyl)amino]-D-glucitol]; abbreviated as PA-0) and a series of specifically modified PA-0 derivatives (Glc m -AltNAc-Glc n -GlcPA, where m + n = 4 and AltNAc is 3-acetoamido-3-deoxy-D-altrose). PA-0 served as an efficient substrate for GP, whereas the other PA-0 derivatives were not as good as the PA-0, indicating that substrate recognition by all the SG1 SG−4 subsites was important for the catalysis of GP. By comparing the initial reaction rate toward the PA-0 derivatives (V derivative) with that toward PA-0 (V PA-0), we found that the value of V derivative/V PA-0 decreased significantly as the level of allosteric activation of GP increased. These results suggest that some conformational changes have taken place in the maltooligosaccharide-binding region of the GP catalytic site during allosteric regulation.


Glycogen Glycogen phosphorylase Modified maltooligosaccharide Pyridylamination Substrate recognition 









2,5-dihydroxybenzoic acid


Glycogen debranching enzyme




α-D-glucose 1-phosphate




Glycogen phosphorylase


High-performance liquid chromatography


Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry


Molecular weight






Inorganic phosphate


Compliance with ethical standards

Conflict of interest

Authors declare no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Chemistry, Graduate School of ScienceOsaka Prefecture UniversityOsakaJapan
  2. 2.Bioorganic Research Institute, Suntory Foundation for Life SciencesKyotoJapan

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