High Affinity Binding of Receptor-associated Protein to Heparin and Low Density Lipoprotein Receptor-related Protein Requires Similar Basic Amino Acid Sequence Motifs

Lora Melman, Zhao Feng Cao, Stephanie Rennke, Maria Paz Marzolo, Mark R. Wardell, Guojun Bu

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

The 39-kDa receptor-associated protein (RAP) is a specialized chaperone for members of the low density lipoprotein receptor gene family, which also binds heparin. Previous studies have identified a triplicate repeat sequence within RAP that appears to exhibit differential functions. Here we generated a series of truncated and site-directed RAP mutants in order to define the sites within RAP that are important for interacting with heparin and low density lipoprotein receptor-related protein (LRP). We found that high affinity binding of RAP to heparin is mediated by the carboxyl-terminal repeat of RAP, whereas both the carboxyl-terminal repeat and a combination of amino and central repeats exhibit high affinity binding to LRP. Several motifs were found to mediate the binding of RAP to heparin, and each contained a cluster of basic amino acids; among them, an intact R282VSR285SR 287EK289 motif is required for high affinity binding of RAP to heparin, whereas two other motifs, R203LR205R 206 and R314ISR317AR319, also contribute to this interaction. We also found that intact motifs of both R 203LR205R206 and R282VSR 285SR287EK289 are required for high affinity binding of RAP to LRP, with the third motif, R314ISR 317AR319, contributing little to RAP-LRP interaction. We conclude that electrostatic interactions likely contribute significantly in the binding of RAP to both heparin and LRP and that high affinity interaction with both heparin and LRP appears to require mostly overlapping sequence motifs within RAP.

Original languageEnglish (US)
Pages (from-to)29338-29346
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number31
DOIs
StatePublished - Aug 3 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'High Affinity Binding of Receptor-associated Protein to Heparin and Low Density Lipoprotein Receptor-related Protein Requires Similar Basic Amino Acid Sequence Motifs'. Together they form a unique fingerprint.

  • Cite this