Sandbox Reserved 433

Introduction

This is a sample scene created with SAT to by Group, and another to make of the protein.

Introduction guide line: Glycogen Synthase Kinase-3 (GSK-3)

1. Begin with "GSK-3 is serine/threonine protein kinase..." explain what that is (structure and general function) 2. Speak about GSK-3 alpha and GSK-3 beta. 3. Begin to describe green screen in relation to the general structure of serine/threonine protein kinase --> specific differences in GSK-3 4.Talk about which species contain GSK-3

 i. Speak about where in which species bodies GSK-3 is present versus humans 
 ii. Explain function in different species structures versus humans 

5. Talk about interactions with other molecules and effects of these interactions.

 i. Effects of changing concentrations of GSK-3 and effects of changing the concentrations of molecules it interacts with 
 ii. Talk about potential effects of mutations 

6. Lead previous section (effects of varying amounts and interactions) into diseases caused by GSK-3. Talk about potential therapeutic uses of it and new treatments under research.

Overall Structure

The overall structure of the Glycogen Synthase Kinase-3β (GSK-β) enzyme is a monomer, and is similar to that of the apoenzyme. GSK-3β has a two-domain kinase fold with a β-strand domain (shown in light blue) at the N-terminus, and an α-helical domain (shown in red) at the C-terminus. Bordering the ATP-binding site is a hinge and glycine-rich loop, found at the boundary of the α-helical and β-strand domain. The activation loop, found along the surface of the substrate-binding groove, can be seen in the (shown in green). The C-terminal resides form a domain that packs against the α -helical domain, which are outside the main kinase fold. The β-strand domain consists of seven β-strands that run in an antiparallel formation. These seven strands include a β -barrel that is interrupted in the middle by a short helix (shown in orange) that packs against the β-barrel. Two of the helixes residues, which are conserved in all kinases, play key roles in the catalytic action of the enzyme. Ser9 is one phosphorylation site that is responsible for the inactivation of GSK-3 β, and the phosphorylation of site Tyr210 is responsible for increasing the catalytic activity. Tyr201 is found on the activation loop, in (green).

Binding Interactions

1)Folding of protein

  a) location and description of position of alpha and beta sheets (Show in separate color on green screen)
  b) crystal structure data (Table 1, Structural Characterizaiton of the GSK-3B Active Site Using Selective and Non-selective ATP-mimetic Inhibitors)

2)Substrate binding

  a)  “phosphate-binding” pocket: describe three crucial basic residues (Show in different color on green screen)
  b)  GSK-3 substrates and binding specifics- S/TxxxS/T (S=Serine, T=threonine, X= any amino acid)

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3217193/ http://www.rcsb.org/pdb/explore/explore.do?pdbId=1Q3D

Additional Features

1. Difference that can be observed from complexes of Staurosporine with GSK-3 beta and other protein kinases such as CDK2, Chk1, Lck and PKA

+ In GSK-3 beta complex with Staurosporine, water is a part of a hydrogen-bonding network

+ Only direct hydrogen bonds are observed between GSK-3 beta and Staurosporine

2. Comparison of GSK-3 beta complexes with Staurosporine and other inhibitors (AMP-PNP, indirubin-3'-monoxime)

+ Between GSK-3 beta complex with Stauroporine and AMP-PNP, the position of N-terminal domain varies.

+ The angle of binding in the active site are different

+ Water is also used as intermediate in hydrogen bonding in GSK-3 beta complex with Indirubin-3'-monoxime (same with complex in Staurosporine )

http://www.sciencedirect.com.silk.library.umass.edu/science/article/pii/S0022283603010593

Quiz Question 1

Which isoform of GSK-3 would work best for ___ function in ___ pathway?

What type of bonding would GSK-3B exhibit with Staurosporine? A picture of the Staurosporine molecule is found below.

The green screen above may help you as well.

Answer: hydrogen bonding

Quiz Question 2

What are the locations of the active sites with respect to the two isoforms?

These green screens may help you. Both scenes are interactive; be sure change perspective and zoom to fully explore the structure. Use the provided color key to interpret the second scene.

Answer: At the interface of the N-terminus beta strand domain and the C-terminus alpha helix domain

J.A. Bertrand, S. Thieffine, A. Vulpetti, C. Cristiani, B. Valsasina, S. Knapp, H.M. Kalisz, M. Flocco, Structural Characterization of the GSK-3β Active Site Using Selective and Non-selective ATP-mimetic Inhibitors, Journal of Molecular Biology, Volume 333, Issue 2, 17 October 2003, Pages 393-407, ISSN 0022-2836, http://dx.doi.org/10.1016/j.jmb.2003.08.031.

(http://www.sciencedirect.com/science/article/pii/S0022283603010593)

See Also

• UvrABC
• Glycogen Synthase Kinase 3
• Serine/Threonie Protein Kinase
• Glycogen Synthase Kinase 3 Beta complexed with Axin Peptide and Inhibitor 7d
• Glycogen Synthase Kinase 3 Beta complexed with AMP-PNP
• Glycogen Synthase Kinase 3 Beta complexed with Axin Peptide and Leucettine L4
• Glycogen Synthase Kinase 3 beta (GSK3B) complexed with a ruthenium octasporine ligand (OS1)

Works Cited

Haar, Ernst ter., Coll, Joyce T., Austen, Douglas A., Hsiao, Hsun-Mei., Swenson, Lora., Jain, Jugnu. Structure of GSK3B reveals a primed phosphorylation mechanism. Vertex Pharmaceuticals Incoprporated, Cambridge MA. Nature Publishing Group, Vol 8 No 7. July 2001.

Credits

Introduction - Zachary Plourde

Overall Structure - Sarah Johnson

Binding Interactions - Christina Lincoln

Additional Features - Bach Pham & Elvan Cevac

Quiz Question 1 - Nerses Haroutunian

Quiz Question 2 - Nick H-K

References