HOMEWORK#5

Protein Data Bank

• due on 4/25

• 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.

 

---

PC Lyu

 

 Answers :

<1>

a) 1EMA

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

c)

• Organism_scientific: Aequorea Victoria
• Organism_common: Jellyfish Plasmid: Prsetb (Invitrogen)
• Expression_system: Escherichia Coli
• Expression_system_strain: Jm109 (De3)
• Other_details: The N-Terminal His-Tag Has Been Removed

d) 708 residues

e) 4 helixs

f)

helix 1 : 1 Gly - 4 Phe

helix 2 : 2 Trp - 57 Leu

helix 3 : 3 Gln - 69 Phe

helix 4 : 4 Phe - 83 Ser

g) 11 b-sheets






<2>

<1EMA>










<1GFL>Chromophore Containing Protein -Ser65, Tyr66, Gly67






<1EMB>








<3>

<1BFP>Mutation: S65, H66, And G67 Are Replaced With Iic 66, Y145F