Homework 5

The Question:

• 1. Please find the 1.9 Å crystal structure of the Aequorea victoria green fluorescent protein (GFP) from PDB. Give the following informations:
• a.) PDB ID number.
• b.) Name of the authors.
• c.) Expression system used by authors.
• d.) Residue number.
• e.) Number of helix.
• f.) The starting residue and ending residue of each helix.
• g.) Number of b-sheet.

• 2. Show three different Rasmol pictures of GFP:
  • whatever you like.
  • highlight the chromophore by spacefill and green.
  • a rotating molecule by GIF builder.

• 3. A blue emission variant of GFP has been made by mutation of two residues. Please find the structure of this blue emission variant. Show its structure and color the two mutated positions blue.

The Answer :

1. (a) 1EMA

    (b) M.Ormo, A.B.Cubitt, K.Kallio, L.A.Gross, R.Y.Tsien, S.J.Remington

    (c) Expression system: Escherichia Coli

    (d) total 236 residue

    (e) 4 helixs

    (f) 1st helix starting residue:gly ending residue:phe

         2nd helix starting residue:trp ending residue:leu

         3re helix starting residue:gln ending residue:phe

         4rd helix starting residue:phe endind residue:ser

   (g) total 11 b-sheets

2.

The green fluorescent protein structure

    we highlight the chromophore by spacefill and green

Please presser this to see rotate molecule

3.Structure of GFP and color the two mutated positions blue. The mutated position is 66 and 145.

  In protein data bank, the PDB id code of blue variant of Green Fluorescent   Protein is 1BFP. The authors of paper is R.M.Wachter, B.A.King, R.Heim, K.Kallio, R.Y.Tsien, S.G.Boxer,S.J.Remington. Expression system is E. coli. MUTATION: S65, H66, AND G67 ARE REPLACED WITH IIC 66, Y145F. [the paper abtract]