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Protein Design

 

Gibas and Subramaniam (1997)

- make bR into a soluble protein

- only computational, no experimental validation

- for validation of such an approach, see Mitra, Steitz, Engelman (2002) Pot. Eng 15, 485-492

- review of protein design approaches:

        Hecht et al., 1990

        Quinn et al., 1994

        Munson et la., 1994

        Lovejoy et al., 1993

        Betz et al., 1995

        Choma et al, 1994

        Robertson et al., 1994

        Bianchi et al., 1994

        Schafmeister et al., 1993

- strategy

    1. Increase Polar Surface Area to make protein soluble

        in membrane proteins: fractional polar surface area of 0.3

        in soluble proteins: 0.5-0.6

    2. replace non-essential amino acids, but keep essential amino acids  [experimentally determined, highly conserved, in contact with retinal, hydrogen bonds, salt bridges, polar residues on surface, Pro, loops]

    3. increase contacts between pairs of helices without filling existing cavities in the protein interior

    4. increase polar residue number without changing charge balance

    5. eliminate amino acids peculiar to membrane proteins, e.g. Trp at helix ends, and Gly

    6. remove amphipathicity of helices

    7. reinforce good capping sequences at the N- and C0termini of helices

 

- evaluation

    homology modeling

 

- Amino acid pairwise preference tables

    determine pairwise preferences of occurrence for the 20 amino acids near N-cap and C-cap based on DSSP definitions