Investigation of Ligand Binding to Low Density Lipoprotein Receptor Related Protein (LRP) PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Investigation of Ligand Binding to Low Density Lipoprotein Receptor Related Protein (LRP) PDF full book. Access full book title Investigation of Ligand Binding to Low Density Lipoprotein Receptor Related Protein (LRP) by Holly Hahn. Download full books in PDF and EPUB format.
Author: Holly Hahn
Publisher:
ISBN:
Category : Carrier proteins
Languages : en
Pages : 110
Get Book Here
Book Description
Author: Holly Hahn
Publisher:
ISBN:
Category : Carrier proteins
Languages : en
Pages : 110
Get Book Here
Book Description
Author: Miklos Guttman
Publisher:
ISBN:
Category :
Languages : en
Pages : 184
Get Book Here
Book Description
The LDLR family of receptors mediates the uptake of lipoprotein particles, and is essential for cholesterol homeostasis. The LDL receptor-related protein 1 (LRP-1) mediates internalization of a large number of diverse ligands and is widely implicated in Alzheimer's disease. Clusters of complement-type ligand binding repeats (CRs) in the LDL receptor family are thought to mediate the interactions between these receptors and their various ligands. Apolipoprotein E, a key ligand for cholesterol homeostasis, has been shown to interact with LDLR, LRP and VLDLR, through these clusters. LDLR and VLDLR each contain a single ligand-binding repeat cluster, whereas LRP contains three large clusters of ligand binding repeats, each with ligand binding functions. In order to study smaller units of these ligand binding clusters we have engineered a new approach to express and refold complement repeat (CR) domains in E. coli. This successfully produced high yields of refolded protein with the benefit of inexpensive isotope labeling for NMR studies. We have expressed a subdomain of sLRP3 (CR16-18) that has previously been shown to recapitulate ligand binding to the isolated receptor binding portion of ApoE (residues 130-149). Binding experiments with the ApoE recognition region of LDLR (LA3-5) and CR16-18 showed that each CR could interact with ApoE(130-149) and that a conserved W25/D30 pair within each repeat appears critical for high affinity. The triple repeat LA3-5 showed the expected interaction with the lipid complexed ApoE(1-191)*DMPC, but surprisingly CR16-18 did not interact with this form of ApoE. To understand these differences in ApoE binding affinity, we introduced mutations of conserved residues from LA5 into CR18, and produced a CR16-18 variant capable of binding ApoE(1-191)*DMPC. This change cannot fully be accounted for by the interaction with ApoE's proposed receptor binding region, therefore we speculate that LA5 is recognizing a distinct epitope on ApoE that may only exist in the lipid bound form. The combination of avidity effects with this distinct recognition process likely governs the ApoE-LDL receptor interaction. Since even the strongest interaction between ApoE(130-149) and a single repeat (CR17) was relatively weak, we constructed a CR17-ApoE(130-149) fusion protein to stabilize the interface for structural studies. The structure revealed a motif seen previously in all ligand CR interactions, in which lysine residues of the ligand interact with the calcium binding site of the CR. Like many ligands of CRs ApoE(130-149) binds as a helix, but with an unexpected turn at H140. These studies also revealed that little structural rearrangement occurs within CR17 upon binding. In addition, dynamics measurements of the free and bound CR17 reveal that certain regions become more ordered, while others become less ordered upon binding. The cytoplasmic tail of LRP, containing two NPXY motifs, has been implicated in the onset of Alzheimer's disease. To examine the intracellular interactions of LRP, as well as to separate which proteins bind to each NPXY motif and their phosphorylation dependence, each NPXY motif microdomain was prepared in both phosphorylated and non-phosphorylated forms and used to probe rodent brain extracts for binding proteins. Proteins that bound specifically to the microdomains were identified by LC-MS/MS, and confirmed by western blot. Recombinant proteins were then tested for binding to each NPXY motif. The NPXY450-- (membrane distal) was found to interact with a large number of proteins, many of which only bound the tyrosine-phosphorylated form. This microdomain also bound a significant number of other proteins in the unphosphorylated state. Many of the interactions were later confirmed to be direct with recombinant proteins. The NPXY44--3 (membrane proximal) bound many fewer proteins and only to the phosphorylated form.
Author: Angela Dawn Williams
Publisher:
ISBN:
Category : Carrier proteins
Languages : en
Pages : 120
Get Book Here
Book Description
Author: Johnny Eugene Croy
Publisher:
ISBN:
Category :
Languages : en
Pages : 390
Get Book Here
Book Description
Author:
Publisher: ScholarlyEditions
ISBN: 1481614037
Category : Science
Languages : en
Pages : 56
Get Book Here
Book Description
Lipoprotein Receptors—Advances in Research and Application: 2012 Edition is a ScholarlyBrief™ that delivers timely, authoritative, comprehensive, and specialized information about Lipoprotein Receptors in a concise format. The editors have built Lipoprotein Receptors—Advances in Research and Application: 2012 Edition on the vast information databases of ScholarlyNews.™ You can expect the information about Lipoprotein Receptors in this eBook to be deeper than what you can access anywhere else, as well as consistently reliable, authoritative, informed, and relevant. The content of Lipoprotein Receptors—Advances in Research and Application: 2012 Edition has been produced by the world’s leading scientists, engineers, analysts, research institutions, and companies. All of the content is from peer-reviewed sources, and all of it is written, assembled, and edited by the editors at ScholarlyEditions™ and available exclusively from us. You now have a source you can cite with authority, confidence, and credibility. More information is available at http://www.ScholarlyEditions.com/.
Author: Richard B. M. Schasfoort
Publisher: Royal Society of Chemistry
ISBN: 1782627308
Category : Science
Languages : en
Pages : 555
Get Book Here
Book Description
Surface plasmon resonance (SPR) plays a dominant role in real-time interaction sensing of biomolecular binding events, this book provides a total system description including optics, fluidics and sensor surfaces for a wide researcher audience.
Author: Neale Ridgway
Publisher: Elsevier
ISBN: 0444634495
Category : Science
Languages : en
Pages : 625
Get Book Here
Book Description
Biochemistry of Lipids: Lipoproteins and Membranes, Volume Six, contains concise chapters that cover a wide spectrum of topics in the field of lipid biochemistry and cell biology. It provides an important bridge between broad-based biochemistry textbooks and more technical research publications, offering cohesive, foundational information. It is a valuable tool for advanced graduate students and researchers who are interested in exploring lipid biology in more detail, and includes overviews of lipid biology in both prokaryotes and eukaryotes, while also providing fundamental background on the subsequent descriptions of fatty acid synthesis, desaturation and elongation, and the pathways that lead the synthesis of complex phospholipids, sphingolipids, and their structural variants. Also covered are sections on how bioactive lipids are involved in cell signaling with an emphasis on disease implications and pathological consequences. Serves as a general reference book for scientists studying lipids, lipoproteins and membranes and as an advanced and up-to-date textbook for teachers and students who are familiar with the basic concepts of lipid biochemistry References from current literature will be included in each chapter to facilitate more in-depth study Key concepts are supported by figures and models to improve reader understanding Chapters provide historical perspective and current analysis of each topic
Author: Norelle L. Daly
Publisher:
ISBN:
Category : Cell receptors
Languages : en
Pages : 360
Get Book Here
Book Description
Author: Rita Kohen Avramoglu
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Get Book Here
Book Description
The low density lipoprotein receptor-related protein (LRP) is a 600 kDa endocytic plasma membrane receptor. LRP is involved in the uptake of numerous structurally and functionally unrelated ligands, including alpha2-macroglobulin:protease complexes (alpha2M*), apolipoprotein E-enriched remnant lipoproteins, Pseudomonas exotoxin A (PEA), and receptor associated protein (RAP). To date, analysis of LRP ligand-binding regions has proven challenging due to its large size and repeating modular structure. Using a recombinant DNA approach, full-length somatic chicken LRP, as well as several deletion mutants, were stably expressed in a mutant CHO-derived cell line deficient in endogenous LRP expression. LRP100, LRP67 and LRP25 encode 100%, 67% and 25% of the protein respectively, with LRP67 and LRP25 encoding large internal deletions but retaining an N-terminal portion, transmembrane region, and cytoplasmic tail. These three mutants were found to possess varying degrees of ligand-binding activity toward alpha 2M*, RAP, and PEA. Generation of this expression system has allowed the mapping of several LRP ligands to the N-terminal portion of the receptor and should allow further study of domains within LRP responsible for its multifunctionality. It was recently postulated that eight spacer regions within LRP containing repeating YWTD motifs adopt beta-propeller conformations. While developing cell, lines expressing refined mutants encoding deletions of putative ligand-binding regions, disruption of the naturally occurring, ordered arrangement of beta-propeller domains was found to have an effect on intracellular trafficking and plasma membrane presentation of mutant receptors. Although highly expressed, several mutants possessing crippled beta-propellers were not detected at the plasma membrane by biotin labeling. These mutants also exhibited delayed or no resistance to endoglycosidase H (endo H) suggesting ER exit was delayed or impaired. Restoration of integral beta-propellers and flanking EGF modules restored ER exit and plasma membrane presentation to a transport incompetent mutant. The beta-propeller domains may play an important role in conferring structural stability to LRP. Caution should therefore be exercised in the mutagenesis of this enormous receptor.
Author: Sangdun Choi
Publisher: Springer
ISBN: 9781493968008
Category : Medical
Languages : en
Pages : 6330
Get Book Here
Book Description
The second edition of this encyclopedia presents over 400 biologically important signaling molecules and the content is built on the core concepts of their functions along with early findings written by some of the world’s foremost experts. The molecules are described by recognized leaders in each molecule. The interactions of these single molecules in signal transduction networks will also be explored. This encyclopedia marks a new era in overview of current cellular signaling molecules for the specialist and the interested non-specialist alike. Currently, there are more than 30,000 genes in human genome. However, not all the proteins encoded by these genes work equally in order to maintain homeostasis. Understanding the important signaling molecules as completely as possible will significantly improve our research-based teaching and scientific capabilities.
