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Molecular architecture and structural basis of allosteric regulation of eukaryotic phosphofructokinases
Author(s) -
Strater Norbert,
Marek Sascha,
Kuettner E. Bartholomeus,
Kloos Marco,
Keim Antje,
Brüser Antje,
Kirchberger Jürgen,
Schöneberg Torsten
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.10-163865
Subject(s) - allosteric regulation , protein subunit , dimer , chemistry , tetramer , stereochemistry , transferase , structural biology , hydrolase , biochemistry , microbiology and biotechnology , biology , enzyme , gene , organic chemistry
Eukaryotic ATP‐dependent 6‐phospho‐fructokinases (Pfks) differ from their bacterial counterparts in a much more complex structural organization and allosteric regulation. Pichia pastoris Pfk (PpPfk) is, with ~1 MDa, the most complex and probably largest eukaryotic Pfk. We have determined the crystal structure of full‐length PpPfk to 3.05 Å resolution in the T state. PpPfk forms a (αβγ) 4 dodec‐amer of D 2 symmetry with dimensions of 161 × 157 × 233 Å mainly via interactions of the α chains. The N‐terminal domains of the a and β chains have folds that are distantly related to glyoxalase I, but the active sites are no longer functional. Interestingly, these domains located at the 2 distal ends of this protein along the long 2‐fold axis form a (aβ) 2 dimer as does the core Pfk domains;however, the domains are swapped across the tetramerization interface. In PpPfk, the unique γ subunit participates in oligomerization of the αβ chains. This modulator protein was acquired from an ancient S‐adenosylmethionine‐dependent methyl‐transferase. The identification of novel ATP binding sites, which do not correspond to the bacterial catalytic or effector binding sites, point to marked structural and functional differences between bacterial and eukaryotic Pfks.—Strater, N., Marek, S., Kuettner, E. B., Kloos, M., Keim, A., Brüser, A., Kirch‐berger, J., Schoneberg, T. Molecular architecture and structural basis of allosteric regulation of eukary‐otic phosphofructokinases. FASEB J. 25, 89–98 (2011). www.fasebj.org