Abstract
Megalin, an approx. 600 kDa transmembrane glycoprotein that acts as multi-ligand transporter, is a member of the low density lipoprotein receptor gene family. Several cysteine-rich repeats, each consisting of about 40 residues, are responsible for the multispecific binding of ligands. The solution structure of the twelfth cysteine-rich ligand-binding repeat with class A motif found in megalin features two short β-strands and two helical turns, yielding the typical fold with a I–III, II–V and IV–VI disulfide bridge connectivity pattern and a calcium coordination site at the C-terminal end. The resulting differences in electrostatic surface potential compared to other ligand-binding modules of this gene family, however, may be responsible for the functional divergence.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- GSH:
-
reduced glutathione
- GSSG:
-
oxidized glutathione
- GST:
-
glutathione S-transferase
- LDL:
-
low density lipoprotein
- LDLR:
-
low density lipoprotein receptor
- LRP:
-
LDLR-related protein
- Meg-A12:
-
megalin cysteine-rich ligand-binding repeat number 12 with class A motif
- NOESY:
-
nuclear Overhauser enhancement spectroscopy
- RAP:
-
receptor associated protein
- RMSD:
-
root-mean-square deviation
- TEV:
-
tobacco etch virus
- TOCSY:
-
total correlation spectroscopy
- VLDL:
-
very low density lipoprotein
References
Atkins AR, Brereton IM, Kroon PA, Lee HT, Smith R (1998) Calcium is essential for the structural integrity of the cysteine-rich, ligand-binding repeat of the low-density lipoprotein receptor. Biochemistry 37:1662–1670
Bieri S, Atkins AR, Lee HT, Winzor DJ, Smith R, Kroon PA (1998) Folding, calcium binding, and structural characterization of a concatemer of the first and second ligand-binding modules of the low-density lipoprotein receptor. Biochemistry 37:10994–11002
Bu G (1998) Receptor-associated protein: a specialized chaperone and antagonist for members of the LDL receptor gene family. Curr Opin Lipidol 9:149–155
Christensen EI, Birn H (2002) Megalin and cubilin: multifunctional endocytic receptors. Nat Rev Mol Cell Biol 3:258–268
Daly NL, Scanlon MJ, Djordjevic JT, Kroon PA, Smith R (1995a) Three-dimensional structure of a cysteine-rich repeat from the low-density lipoprotein receptor. Proc Natl Acad Sci USA 92:6334–6338
Daly NL, Djordjevic JT, Kroon PA, Smith R (1995b) Three-dimensional structure of the second cysteine-rich repeat from the human low-density lipoprotein receptor. Biochemistry 34:14474–14481
Dolmer K, Huang W, Gettins PGW (2000) NMR solution structure of complement-like repeat CR3 from the low density lipoprotein receptor-related protein. Evidence for specific binding to the receptor binding domain of human α2-macroglobulin. J Biol Chem 275:3264–3269
Fass D, Blacklow S, Kim PS, Berger JM (1997) Molecular basis of familial hypercholesterolaemia from structure of LDL receptor module. Nature 388:691–693
Huang W, Dolmer K, Gettins PGW (1999) NMR solution structure of complement-like repeat CR8 from the low density lipoprotein receptor-related protein. J Biol Chem 274:14130–14136
Johnson BA, Blevins RA (1994) NMR View: a computer program for the visualization and analysis of NMR data. J Biomol NMR 4:603–614
Laskowski RA, Rullmann JA, MacArthur MW, Kaptein R, Thornton JM (1996) AQUA and PROCHECK-NMR: programs for checking the quality of protein structures solved by NMR. J Biomol NMR 8:477–486
Linge JP, Habeck M, Rieping W, Nilges M (2003) ARIA: automated NOE assignment and NMR structure calculation. Bioinformatics 19:315–316
Melcher K (2000) A modular set of prokaryotic and eukaryotic expression vectors. Anal Biochem 277:109–120
Nicholls A, Sharp KA, Honig B (1991) Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins 11:281–296
North CL, Blacklow SC (2000) Solution structure of the sixth LDL-A module of the LDL receptor. Biochemistry 39:2564–2571
Russell DW, Brown MS, Goldstein JL (1989) Different combinations of cysteine-rich repeats mediate binding of low density lipoprotein receptor to two different proteins. J Biol Chem 264:21682–21688
Saito A, Yamazaki H, Rader K, Nakatani A, Ullrich R, Kerjaschki D, Orlando RA, Farquar MG (1996) Mapping rat megalin: the second cluster of ligand binding repeats contains a 46-amino acid pathogenic epitope involved in the formation of immune deposits in Heymann nephritis. Proc Natl Acad Sci USA 93:8601–8605
Simonovic M, Dolmer K, Huang W, Strickland DK, Voltz K, Gettins PGW (2001) Calcium coordination and pH dependence of the calcium affinity of ligand-binding repeat CR7 from the LRP. Comparison with related domains from the LRP and the LDL receptor. Biochemistry 40:15127–15134
Wishart DS, Bigam CG, Yao J, Abildgaard F, Dyson HJ, Oldfield E, Markley JL, Sykes BD (1995) 1H, 13C and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR 6:135–140
Wüthrich K (1986) NMR of proteins and nucleic acids. Wiley, New York
Acknowledgements
The authors would like to thank Sigurd Krieger (Medical University of Vienna) for kindly providing the cDNA cassette of Meg-A12, Dr. Frank Bernhard (University of Frankfurt) for kindly providing (His)7-TEV-protease, and Dr. Vladimir Rogov for helpful discussions. The European Large Scale Facility for Biomolecular NMR at the University of Frankfurt is acknowledged for the use of its equipment.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wolf, C.A., Dancea, F., Shi, M. et al. Solution structure of the twelfth cysteine-rich ligand-binding repeat in rat megalin. J Biomol NMR 37, 321–328 (2007). https://doi.org/10.1007/s10858-006-9129-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10858-006-9129-3