Small Protein

structure service
Cryo-electron microscopy (cryo-EM) is a form of transmission electron
microscopy (TEM) where biological samples are studied at cryogenic
temperatures. It is becoming a more and more important technique in
the field of structural biology, as has been manifested by many once
hard-to-determine macromolecular structures. With the rapid
advancement of both hardware and software, the resolution of cryo-
EM maps is improving steadily. In some cases, near-atomic resolution
had been obtained, including those of viruses, ribosomes,
mitochondria, ion channels, and enzyme complexes as small as 150
kD.
However, very small proteins remain challenging to study using EM
because of the difficulty in their visualization, alignment and
validation. To address this technical challenge, Creative Biostructure is
devoted to optimizing our EM protocol and making use of available
resources to provide high-resolution imaging and three-dimensional
reconstruction services for a broad spectrum of small proteins of
interest. Our strategies include:
•Improved sample preparation with ultra-thin vitreous ice layer
•Selection of binding agents such as Fabs
•Optimization of the electron dose
•Variation of defocus
•Energy filter and correction of beam-induced motion
•Use of phase plate for contrast enhancement
The advantages of using our EM platform for the structure determination
of proteins with small molecular weight include sample-saving, fast
turnaround time, and cost-effective. Creative Biostructure promises to
work closely with our customers to provide tailored cryo-EM strategies for
determining the structure of biologically or structurally important small
proteins, with a resolution comparable to traditional X-ray crystallography
techniques.
References:
•Merk A, et al. (2016) “Breaking cryo-EM resolution barriers to facilitate
drug discovery”. Cell165(7):1698-1707.
•Wu S, et al. (2012) “Fabs enable single particle cryo-EM studies of small
proteins”. Structure20(4):582-592.
Excerpt from:
https://www.creative-biostructure.com/cryo-em-for-small-
proteins-561.htm