Stereospecific assignments of glycine in proteins by stereospecific deuteration and {sup 15}N labeling

Conference · Thu Dec 01 04:00:00 EST 1994

OSTI ID:83397

Hansen, A P; Curley, Jr, R W; Panigot, M J; Fesik, S W ^[1]

---

1. Ohio State Univ., Columbus, OH (United States)

Stereospecific assignments are important for accurately determining the three-dimensional structures of proteins through the use of multidimensional NMR techniques. It is especially important to stereospecifically assign the glycine {alpha}-protons in proteins because of the potential for different backbone conformations of this residue. These stereospecific assignments are critical for interpreting the {sup 3}J{sub NH,{alpha}H} coupling constants and NOEs involving the glycine {alpha}-protons that determine the conformation of this part of the protein. However, it is often difficult to unambiguously obtain the stereospecific assignments for glycine residues by using only NOE data. In this poster, we present a method for unambiguous, stereospecific assignment of the {alpha}-protons of glycine residues. This method involves synthesis of stereo-specifically deuterated and {sup 15}N-labeled Gly using a slightly modified procedure originally described by Woodard and coworkers for the stereoselective deuteration of glycine. The stereospecifically deuterated and {sup 15}N-labeled Gy has been incorporated into recombinant proteins expressed in both bacterial systems (FKBP) and mammalian cells (u-PA). Two- and three-dimensional isotope-filtered and isotope-edited NMR experiments were used to obtain the stereospecific assignments of the glycine {alpha}-protons for these proteins.

• Research Organization: Los Alamos National Lab., NM (United States)
• OSTI ID: 83397
• Report Number(s): LA--12893-C; CONF-9403228--; ON: DE95012795
• Country of Publication: United States
• Language: English

Related Subjects

40 CHEMISTRY
55 BIOLOGY AND MEDICINE
BASIC STUDIES
COUPLING CONSTANTS
DEUTERATION
GLYCINE
ISOTOPE APPLICATIONS
NITROGEN 15
NMR IMAGING
NUCLEAR MAGNETIC RESONANCE
PROTEIN STRUCTURE
PROTEINS
STEREOCHEMISTRY