NMR structural studies of domain 1 of receptor-associated protein.
The 39 kDa receptor-associated protein (RAP) is an endoplasmic reticulum resident protein that binds tightly to the low-density lipoprotein receptor-related protein (LRP) as well as to other members of the low-density lipoprotein receptor superfamily. The association of RAP with LRP prevents this receptor from interacting with ligands. RAP is a three-domain protein that contains two independent LRP binding sites; one located within domains 1 and 2, and one located within domain 3. As the first step toward defining the structure of the full-length protein and understanding the interaction between RAP and this family of receptors, we have determined the 3D structure of domain 1 using constraints derived from heteronuclear multi-dimensional NMR spectra, including NOEs, dihedral angles, J-couplings and chemical shifts, as well as two sets of non-correlated residual dipolar couplings measured from the protein solutions in anisotropic media of Pf1 and 6% polyacrylamide gel. The backbone C(alpha) rmsd between the current structure and a homo-nuclear NOE-based structure is about 2 A. The large rmsd mainly reflects the significant differences in helical orientation and in the structural details of the long helix (helix 2) between the two structures.
Wu
YiBing
Y
Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, National Cancer Institute at Frederick, National Institutes of Health, Frederick, MD 21702, USA.
Migliorini
Molly
M
Walsh
Joseph
J
Yu
Ping
P
Strickland
Dudley K
DK
Wang
Yun-Xing
YX
D000180:Acrylic Resins / Q000737:chemistry; D016880:Anisotropy; D001665:Binding Sites; D002244:Carbon / Q000737:chemistry; D019992:Databases as Topic; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D006860:Hydrogen Bonding; D026502:LDL-Receptor Related Proteins / Q000737:chemistry*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D011522:Protons; D012984:Software; D000099166:Polyacrylamides
D000180:Acrylic Resins; D002244:Carbon; D026502:LDL-Receptor Related Proteins; D008024:Ligands; D011506:Proteins; D011522:Protons; D000099166:Polyacrylamides