CRYOPROBE FOR A 600 MHZ NMR SPECTROMETER

Harvard University (Medical School), Boston MA

Investigators

Linked publications & trials

Abstract

This is a proposal to purchase a cryoprobe for a 600 MHz NMR instrument. This probe will be shared between four principal investigators at Harvard University, Gerhard Wagner, Gregory Verdine, Stephen Blacklow and Ellis Reinherz. This cryoprobe will enhance the sensitivity of our 600 MHz spectrometer three- to four-fold. This is dramatic considering that the sensitivity gain from 600 MHz to 750 MHz is only about 1.5. To achieve fourfold gain in signal to noise by signal averaging one would have to acquire sixteen times the number of scans. The cryoprobe will thus enable us to determine structures of proteins and protein complexes that are only soluble up to 0.1 mM concentration. The second important aspect of a cryoprobe at a 600 MHz spectrometer is that it will be at the optimal field strength for using the TROSY on aromatic CH groups. NOES to aromatic side chains form the most important source of long-range NOES for NMR structure determination. The cryoprobe will be used by Gerhard Wagner to solve large complexes of proteins involved in translation initiation and in apoptosis. The groups of Ellis Reinherz and Gerhard Wagner will collaborate to solve structures of a single-chain CD3epsilon gamma and its interaction with a TCR. Furthermore, they will work on a structure of CD3epsilon delta and its interaction with CD4. The groups of Gregory Verdine and Gerhard Wagner will collaborate on solving a complex of the DNA- repair protein ada with DNA. The group of Stephen Blacklow will work on domains of the LDL receptor and the LIN-12 domain of human Notch 1.