3D NMR experiments for measuring 15N relaxation data of large proteins: application to the 44 kDa ectodomain of SIV gp41.
A suite of 3D NMR experiments for measuring 15N-¿1H¿ NOE, 15N T1, and 15N T1rho values in large proteins, uniformly labeled with 15N and 13C, is presented. These experiments are designed for proteins that exhibit extensive spectral overlap in the 2D 1H-15N HSQC spectrum. The pulse sequences are readily applicable to perdeuterated samples, which increases the spectral resolution and signal-to-noise ratio, thereby permitting the characterization of protein dynamics to be extended to larger protein systems. Application of the pulse sequences is demonstrated on a perdeuterated 13C/15N-labeled sample of the 44 kDa ectodomain of SIV gp41.
Caffrey
M
M
National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, 20892-0520, USA.
Kaufman
J
J
Stahl
S J
SJ
Wingfield
P T
PT
Gronenborn
A M
AM
Clore
G M
GM
D002247:Carbon Isotopes; D003903:Deuterium; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008562:Membrane Glycoproteins / Q000032:analysis*; D008970:Molecular Weight; D009587:Nitrogen Isotopes; D011487:Protein Conformation; D012191:Retroviridae Proteins / Q000032:analysis*; D012815:Signal Processing, Computer-Assisted; D015302:Simian Immunodeficiency Virus / Q000737:chemistry*; D014759:Viral Envelope Proteins / Q000032:analysis*
D002247:Carbon Isotopes; D008562:Membrane Glycoproteins; D009587:Nitrogen Isotopes; D012191:Retroviridae Proteins; C082614:SIV envelope protein gp41; D014759:Viral Envelope Proteins; D006859:Hydrogen; D003903:Deuterium