NMR Structure of Sec35p/Cog2p
EMSL Project ID
2592
Abstract
We are requesting NMR time to perform experiments aimed at determining the three-dimensional structure of yeast Sec35p/Cog2p. This 32 kD protein is an essential component of the yeast Conserved Oligomeric Golgi (COG) complex important for intracellular transport. The sequence of Sec35p/Cog2p bears no resemblance to that of any structurally characterized protein, suggesting that it may adopt a novel fold. Furthermore, no structural information is currently available for any COG complex subunit. We have identified a 23 kD fragment of Sec35p that is highly soluble, monomeric, and appears to contain all of the regular secondary structural elements present in the full-length polypeptide. 15N- and 13C,15N-labeled NMR samples at 1 mM protein concentrations have been produced and are stable for many weeks. HSQC spectra collected using a 600 MHz spectrometer show reasonable peak dispersion and provide preliminary evidence that a full structure determination by NMR is feasible. Lorraine Cavanaugh, the senior graduate student carrying out this work, has received and will continue to receive continuous assistance from our collaborator Jose Rizo-Rey (UT Southwestern), a distinguished NMR spectroscopist, during the course of this structure determination. We anticipate that the structure of Sec35p/Cog2p will help elucidate molecular mechanisms of intracellular transport, a fundamental problem in cell biology.
Project Details
Project type
Capability Research
Start Date
2002-10-01
End Date
2003-04-24
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Cavanaugh L, X Chen, B Richardson, D Ungar, I Pelczer, J Rizo, and FM Hughson. 2007. "Structural Analysis of Conserved Oligomeric Golgi Complex Subunit 2." Journal of Biological Chemistry 282(32):23418-23426. doi:10.1074/jbc.M703716200