TY - JOUR
T1 - Protein production from the structural genomics perspective
T2 - Achievements and future needs
AU - Almo, Steven C.
AU - Garforth, Scott J.
AU - Hillerich, Brandan S.
AU - Love, James D.
AU - Seidel, Ronald D.
AU - Burley, Stephen K.
N1 - Funding Information:
This work was supported by NIH grants U01GM094665 , U54GM094662 , U54GM093342 , and P30CA013330 .
Funding Information:
Other structural genomics centers are implementing high throughput eukaryotic expression pipelines. Courtesy of the Genomics Institute of the Novartis Research Foundation (GNF), the Joint Center for Structural Genomics (JCSG) has access to the highly automated Protein Expression and Purification Platform (PEPP), a fully automate robotic solution to high-throughput small-scale expression evaluation in eukaryotic cells [ 54 ]. The Structural Genomics Consortium (SGC) has established a somewhat less automated platform that enables throughputs of up to 250 L of baculovirus-infected insect cells/month, supporting work on a host of challenging human targets, including tyrosine and serine/threonine kinases and integral membrane proteins ( http://www.thesgc.org ). These large-scale efforts are complemented by a broad array of technology development programs supported by both the NIH and international research funding agencies (e.g. see http://grants.nih.gov/grants/guide/pa-files/PAR-13-032.html ).
PY - 2013/6
Y1 - 2013/6
N2 - Despite a multitude of recent technical breakthroughs speeding high-resolution structural analysis of biological macromolecules, production of sufficient quantities of well-behaved, active protein continues to represent the rate-limiting step in many structure determination efforts. These challenges are only amplified when considered in the context of ongoing structural genomics efforts, which are now contending with multi-domain eukaryotic proteins, secreted proteins, and ever-larger macromolecular assemblies. Exciting new developments in eukaryotic expression platforms, including insect and mammalian-based systems, promise enhanced opportunities for structural approaches to some of the most important biological problems. Development and implementation of automated eukaryotic expression techniques promises to significantly improve production of materials for structural, functional, and biomedical research applications.
AB - Despite a multitude of recent technical breakthroughs speeding high-resolution structural analysis of biological macromolecules, production of sufficient quantities of well-behaved, active protein continues to represent the rate-limiting step in many structure determination efforts. These challenges are only amplified when considered in the context of ongoing structural genomics efforts, which are now contending with multi-domain eukaryotic proteins, secreted proteins, and ever-larger macromolecular assemblies. Exciting new developments in eukaryotic expression platforms, including insect and mammalian-based systems, promise enhanced opportunities for structural approaches to some of the most important biological problems. Development and implementation of automated eukaryotic expression techniques promises to significantly improve production of materials for structural, functional, and biomedical research applications.
UR - http://www.scopus.com/inward/record.url?scp=84879185251&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84879185251&partnerID=8YFLogxK
U2 - 10.1016/j.sbi.2013.02.014
DO - 10.1016/j.sbi.2013.02.014
M3 - Review article
C2 - 23642905
AN - SCOPUS:84879185251
SN - 0959-440X
VL - 23
SP - 335
EP - 344
JO - Current Opinion in Structural Biology
JF - Current Opinion in Structural Biology
IS - 3
ER -