This is the most common method that is
used to generate the 3D structures of proteins. Predicting the 3D structure
from the amino acid sequence is very difficult so it is almost always measured
experimentally. There are two main aspects

1)    How
do you generate the purified protein in the correct 3D conformation and
crystallise it, and then

We Will Write a Custom Essay Specifically
For You For Only $13.90/page!


order now

2)    How
do your x-ray diffraction experiments give you the structure.

There are important examples in
membrane proteins, and comparisons to the highest resolution structures from
EM.

X-ray crystallography is a technique to
generate the three-dimensional structures of proteins at an atomic level and
understand their functions from its crystallised form. There are three main components
needed to complete an x-ray crystallography analysis – a purified sample at
high concentration which is crystallised, an X-ray source and a detector where
the resulting diffraction patterns are processed.

Why use X-rays?

The
resolution of an image in any form of microscopy depends on the wavelength of
electromagnetic radiation used. Light microscopy (where the range of
wavelengths is 380-750nm) is used to see individual cells and sub-cellular
organelles. Electron microscopy (where the wavelength is approximately 10nm) is
used to see cellular architecture and the shapes of large protein molecules. Therefore,
to see proteins at an atomic level, electromagnetic radiation of around 0.1 nm
should be used – this corresponds to the wavelength of X-rays. 1

Why do we need a crystal?

The diffraction
from a single molecule is too weak to be measured therefore the proteins are
crystallised. The diffraction of a crystal is measurable as there are multiple
copies within them. Typical protein cells crystals are about 0.2mm in size but
usable crystals have been reported from tens of microns to a few millimetres.

2

x

Hi!
I'm Erica!

Would you like to get a custom essay? How about receiving a customized one?

Check it out