Investigation of the interaction between the large and small subunits of potato ADP-glucose pyrophosphorylase.
ADP-glucose pyrophosphorylase (AGPase), a key allosteric enzyme involved in higher plant starch biosynthesis, is composed of pairs of large (LS) and small subunits (SS). Current evidence indicates that the two subunit types play distinct roles in enzyme function. Recently the heterotetrameric structure of potato AGPase has been modeled. In the current study, we have applied the molecular mechanics generalized born surface area (MM-GBSA) method and identified critical amino acids of the potato AGPase LS and SS subunits that interact with each other during the native heterotetrameric structure formation. We have further shown the role of the LS amino acids in subunit-subunit interaction by yeast two-hybrid, bacterial complementation assay and native gel. Comparison of the computational results with the experiments has indicated that the backbone energy contribution (rather than the side chain energies) of the interface residues is more important in identifying critical residues. We have found that lateral interaction of the LS-SS is much stronger than the longitudinal one, and it is mainly mediated by hydrophobic interactions. This study will not only enhance our understanding of the interaction between the SS and the LS of AGPase, but will also enable us to engineer proteins to obtain better assembled variants of AGPase which can be used for the improvement of plant yield.
Baris
Ibrahim
I
Department of Chemical and Biological Engineering, College of Engineering, Koc University, Rumeli Feneri Yolu, Istanbul, Turkey.
Tuncel
Aytug
A
Ozber
Natali
N
Keskin
Ozlem
O
Kavakli
Ibrahim Halil
IH
D000595:Amino Acid Sequence; D019295:Computational Biology / Q000379:methods*; D005816:Genetic Complementation Test; D051599:Glucose-1-Phosphate Adenylyltransferase / Q000235:genetics / Q000378:metabolism*; D006860:Hydrogen Bonding; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D055503:Protein Multimerization; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D011198:Solanum tuberosum / Q000235:genetics*; D013816:Thermodynamics; D020798:Two-Hybrid System Techniques; D014867:Water / Q000378:metabolism
D021122:Protein Subunits; D014867:Water; D051599:Glucose-1-Phosphate Adenylyltransferase