Last class Enzyme inhibition types Inhibitors and kinetic parameters Paper 2.

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Presentation transcript:

Last class Enzyme inhibition types Inhibitors and kinetic parameters Paper 2

Crystallization VS Precipitation

Why crystallize proteins? chemwiki.ucdavis.edu

Protein crystallization

Protein crystallization

Protein crystallization

Sensitivity of protein crystallization

How to get crystals NOT Ppt? Start off with undersaturated Very slowly move to supersaturated Equilibrate to allow nucleation & crystallization Every protein different! Art VS Science!

Protein crystallization

Factors affecting protein crystallization pH

Vaporization to equilibrate

Lysozyme facts Second protein structure First protein sequenced with all 20 amino acids First enzyme mechanism

Lysozyme in eggs…

…and body fluids (eg. mucous)

Lysozyme is defense mechanism PLoS ONE 9(2): e doi: /journal.pone

Breaks bonds in peptidoglycan layer

G+ve VS G-ve

Lab Make egg extracts (get the egg white!) Total protein in egg extract Enzyme activity (OD of Micrococcus luteus ) Set up crystallization trays with purified lysozyme & egg extract