notes

1.  Hi, my name is Ben Mair and I chose to research protein engineering
 To understand why protein engineering is important, we first need to know what proteins
do
 Proteins are found in living organisms
 Important as enzymes
 Biological catalysts
 Organisms would struggle to control destructively harsh reaction conditions
 Chemists look for more sustainable reactions
 Adapting proteins since 70s
 To engineer a protein, we need to know what it is
 Amino acids condensed into polymer
 3D structure due to hydrogen bonding, hydrophilic & hydrophobic interactions, dipole
interactions and disulphide bridges (cysteine)
 Haem prosthetic group (organometallic)
 Proteins 1000s of amino acids long
 Need to look at how nature makes them
 Polymer of nucleotides
 Complementary
 3 nucleotides per protein
 Adenine, thymine and guanine for methionine (start)
Forward 2
 This is all very complex
 Rational almost impossible completely by deduction
 Let nature do it
 Breeding dogs (best function)
 How do you breed proteins?
Back 1
 Endonuclease hydrolyses at GAAT (catalogues)
 DNA polymerase fuses new combination (mutant)
 Sample not large enough
Forward 2
 PCR utilises complementary nature of DNA
 DNA polymerase needs somewhere to attach (sticky end)
 Generic short strands hydrogen bond
 Grow exponentially
 Push for diversity and function difficult
 Rationalise irrational protein design

2.  Map interaction e.g. 2 angstroms
 2D contact matrix
 Change endonuclease
 Proven effective
 An example
 P450 very important
 Oxidises fatty acids
 Increase in molecules as small as propane
 How does this work?
 More complementary active site
 Substrate displaces water
 Higher reduction potential
 Gains electron to form superoxide with oxygen (oxidising)
 However, is this applicable to industry
 Not really
 Struggle to transcribe out of cell
 Not economical
 Recent research into artificial cofactors
 Promising field
 Caltech
 Solving problems, therefore good