Thermus thermophilus Glycoside Hydrolase Family 57 Branching Enzyme: Crystal Structure, Mechanism of Action, and Products FormedPalomo, M., Pijning, T., Booiman, T., Dobruchowska, J. M., Vlist, J. V. D., Kralj, S., Planas, A., Loos, K., Kamerling, J. P., Dijkstra, B. W., Maarel, M. J. E. C. V. D., Dijkhuizen, L. & Leemhuis, H., 4-Feb-2011, In : The Journal of Biological Chemistry. 286, 5, p. 3520-3530 11 p.
Research output: Contribution to journal › Article › Academic › peer-review
Branching enzyme (EC 220.127.116.11; glycogen branching enzyme; GBE) catalyzes the formation of alpha 1,6-branching points in glycogen. Until recently it was believed that all GBEs belong to glycoside hydrolase family 13 (GH13). Here we describe the cloning and expression of the Thermus thermophilus family GH57-type GBE and report its biochemical properties and crystal structure at 1.35-angstrom resolution. The enzyme has a central (beta/alpha)(7)-fold catalytic domain A with an inserted domain B between beta 2 and alpha 5 and an alpha-helix-rich C-terminal domain, which is shown to be essential for substrate binding and catalysis. A maltotriose was modeled in the active site of the enzyme which suggests that there is insufficient space for simultaneously binding of donor and acceptor substrates, and that the donor substrate must be cleaved before acceptor substrate can bind. The biochemical assessment showed that the GH57 GBE possesses about 4% hydrolytic activity with amylose and in vitro forms a glucan product with a novel fine structure, demonstrating that the GH57 GBE is clearly different from the GH13 GBEs characterized to date.
|Number of pages||11|
|Journal||The Journal of Biological Chemistry|
|Publication status||Published - 4-Feb-2011|
- THERMOCOCCUS-LITORALIS, BACTERIAL GLYCOGEN, 4-ALPHA-GLUCANOTRANSFERASE, IDENTIFICATION, PHOSPHORYLASE, EVOLUTION, PROTEINS, MARKER