FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W4313454417> ?p ?o ?g. }

• W4313454417 endingPage "101662" @default.
• W4313454417 startingPage "101662" @default.
• W4313454417 abstract "The liver-derived circulating PCSK9 enhances the degradation of the LDL receptor (LDLR) in endosomes/lysosomes. PCSK9 inhibition or silencing is presently used in clinics worldwide to reduce LDL-cholesterol, resulting in lower incidence of cardiovascular disease and possibly cancer/metastasis. The mechanism by which the PCSK9-LDLR complex is sorted to degradation compartments is not fully understood. We previously suggested that out of the three M1, M2 and M3 subdomains of the C-terminal Cys/His-rich-domain (CHRD) of PCSK9, only M2 is critical for the activity of extracellular of PCSK9 on cell surface LDLR. This likely implicates the binding of M2 to an unknown membrane-associated “protein X” that would escort the complex to endosomes/lysosomes for degradation. We reported that a nanobody P1.40 binds the M1 and M3 domains of the CHRD and inhibits the function of PCSK9. It was also reported that the cytosolic adenylyl cyclase-associated protein 1 (CAP1) could bind M1 and M3 subdomains and enhance the activity of PCSK9. In this study, we determined the 3-dimensional structure of the CHRD-P1.40 complex to understand the intricate interplay between P1.40, CAP1 and PCSK9 and how they regulate LDLR degradation. X-ray diffraction of the CHRD-P1.40 complex was analyzed with a 2.2 Å resolution. The affinity and interaction of PCSK9 or CHRD with P1.40 or CAP1 was analyzed by atomic modeling, site-directed mutagenesis, bio-layer interferometry, expression in hepatic cell lines and immunocytochemistry to monitor LDLR degradation. The CHRD-P1.40 interaction was further analyzed by deep mutational scanning and binding assays to validate the role of predicted critical residues. Conformational changes and atomic models were obtained by small angle X-ray scattering (SAXS). We demonstrate that PCSK9 exists in a closed or open conformation and that P1.40 favors the latter by binding key residues in the M1 and M3 subdomains of the CHRD. Our data show that CAP1 is well secreted by hepatic cells and binds extracellular PCSK9 at distinct residues in the M1 and M3 modules and in the acidic prodomain. CAP1 stabilizes the closed conformation of PCSK9 and prevents P1.40 binding. However, CAP1 siRNA only partially inhibited PCSK9 activity on the LDLR. By modeling the previously reported interaction between M2 and an R-X-E motif in HLA-C, we identified Glu567 and Arg549 as critical M2 residues binding HLA-C. Amazingly, these two residues are also required for the PCSK9-induced LDLR degradation. The present study reveals that CAP1 enhances the function of PCSK9, likely by twisting the protein into a closed configuration that exposes the M2 subdomain needed for targeting the PCSK9-LDLR complex to degradation compartments. We hypothesize that “protein X”, which is expected to guide the LDLR-PCSK9-CAP1 complex to these compartments after endocytosis into clathrin-coated vesicles, is HLA-C or a similar MHC-I family member. This conclusion is supported by the PCSK9 natural loss-of-function Q554E and gain-of-function H553R M2 variants, whose consequences are anticipated by our modeling." @default.
• W4313454417 created "2023-01-06" @default.
• W4313454417 creator A5001789482 @default.
• W4313454417 creator A5007175703 @default.
• W4313454417 creator A5009368039 @default.
• W4313454417 creator A5042563402 @default.
• W4313454417 creator A5043003865 @default.
• W4313454417 creator A5051298126 @default.
• W4313454417 creator A5053379768 @default.
• W4313454417 creator A5065711128 @default.
• W4313454417 creator A5071656424 @default.
• W4313454417 creator A5073145975 @default.
• W4313454417 creator A5075033941 @default.
• W4313454417 creator A5075538723 @default.
• W4313454417 creator A5078178131 @default.
• W4313454417 creator A5080359448 @default.
• W4313454417 creator A5082669251 @default.
• W4313454417 creator A5085774208 @default.
• W4313454417 date "2023-01-01" @default.
• W4313454417 modified "2023-10-18" @default.
• W4313454417 title "Molecular interactions of PCSK9 with an inhibitory nanobody, CAP1 and HLA-C: Functional regulation of LDLR levels" @default.
• W4313454417 cites W1491374834 @default.
• W4313454417 cites W1861627737 @default.
• W4313454417 cites W1879505420 @default.
• W4313454417 cites W1969950216 @default.
• W4313454417 cites W1974955986 @default.
• W4313454417 cites W1980374511 @default.
• W4313454417 cites W1981974356 @default.
• W4313454417 cites W1983029570 @default.
• W4313454417 cites W1986040042 @default.
• W4313454417 cites W1986191025 @default.
• W4313454417 cites W1989660004 @default.
• W4313454417 cites W2009681469 @default.
• W4313454417 cites W2010689685 @default.
• W4313454417 cites W2015860855 @default.
• W4313454417 cites W2038614269 @default.
• W4313454417 cites W2040612932 @default.
• W4313454417 cites W2045315353 @default.
• W4313454417 cites W2047861227 @default.
• W4313454417 cites W2050348933 @default.
• W4313454417 cites W2055571770 @default.
• W4313454417 cites W2056411372 @default.
• W4313454417 cites W2062062360 @default.
• W4313454417 cites W2064994526 @default.
• W4313454417 cites W2069993714 @default.
• W4313454417 cites W2071154679 @default.
• W4313454417 cites W2071885555 @default.
• W4313454417 cites W2076648603 @default.
• W4313454417 cites W2078669364 @default.
• W4313454417 cites W2080023404 @default.
• W4313454417 cites W2082817623 @default.
• W4313454417 cites W2099136586 @default.
• W4313454417 cites W2099492906 @default.
• W4313454417 cites W2099654648 @default.
• W4313454417 cites W2105119801 @default.
• W4313454417 cites W2108921801 @default.
• W4313454417 cites W2110124859 @default.
• W4313454417 cites W2113149246 @default.
• W4313454417 cites W2113577386 @default.
• W4313454417 cites W2117011155 @default.
• W4313454417 cites W2118242699 @default.
• W4313454417 cites W2122864632 @default.
• W4313454417 cites W2135101596 @default.
• W4313454417 cites W2136034547 @default.
• W4313454417 cites W2146660254 @default.
• W4313454417 cites W2148858610 @default.
• W4313454417 cites W2149994005 @default.
• W4313454417 cites W2153255725 @default.
• W4313454417 cites W2158266834 @default.
• W4313454417 cites W2411116279 @default.
• W4313454417 cites W2518792780 @default.
• W4313454417 cites W2551904109 @default.
• W4313454417 cites W2560062929 @default.
• W4313454417 cites W2561255627 @default.
• W4313454417 cites W2566580306 @default.
• W4313454417 cites W2770394202 @default.
• W4313454417 cites W2803368411 @default.
• W4313454417 cites W2811000073 @default.
• W4313454417 cites W2884109240 @default.
• W4313454417 cites W2908533470 @default.
• W4313454417 cites W2911014938 @default.
• W4313454417 cites W2971968715 @default.
• W4313454417 cites W2981286566 @default.
• W4313454417 cites W3103804121 @default.
• W4313454417 cites W3201695038 @default.
• W4313454417 cites W3205769256 @default.
• W4313454417 cites W4210355224 @default.
• W4313454417 cites W4247043122 @default.
• W4313454417 cites W4280501338 @default.
• W4313454417 cites W856782376 @default.
• W4313454417 cites W2910765234 @default.
• W4313454417 doi "https://doi.org/10.1016/j.molmet.2022.101662" @default.
• W4313454417 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36566984" @default.
• W4313454417 hasPublicationYear "2023" @default.
• W4313454417 type Work @default.
• W4313454417 citedByCount "1" @default.
• W4313454417 countsByYear W43134544172023 @default.
• W4313454417 crossrefType "journal-article" @default.

• next