The structure-function relationship of a signaling-competent, dimeric Reelin fragment

Liam S Turk; Xuyuan Kuang; Valentina Dal Pozzo; Khush Patel; Muyuan Chen; Kevin Huynh; Michael J Currie; Daniel Mitchell; Renwick C J Dobson; Gabriella D'Arcangelo; Wei Dai; Davide Comoletti

doi:10.1016/j.str.2021.05.012

The structure-function relationship of a signaling-competent, dimeric Reelin fragment

Structure. 2021 Oct 7;29(10):1156-1170.e6. doi: 10.1016/j.str.2021.05.012. Epub 2021 Jun 4.

Authors

Affiliations

¹ Child Health Institute of New Jersey, New Brunswick, NJ 08901, USA; Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA; School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand.
² Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA; Department of Hyperbaric Oxygen, Central South University, Changsha, Hunan Province, China.
³ Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA.
⁴ Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
⁵ Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA.
⁶ Biomolecular Interactions Centre and School of Biological Sciences, University of Canterbury, Christchurch 8041, New Zealand.
⁷ School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand.
⁸ Biomolecular Interactions Centre and School of Biological Sciences, University of Canterbury, Christchurch 8041, New Zealand; Bio21 Molecular Science and Biotechnology Institute, Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC 3010, Australia.
⁹ Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA. Electronic address: wei.dai@rutgers.edu.
¹⁰ Child Health Institute of New Jersey, New Brunswick, NJ 08901, USA; Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA; School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand. Electronic address: davide.comoletti@vuw.ac.nz.

PMID: 34089653
DOI: 10.1016/j.str.2021.05.012

Abstract

Reelin operates through canonical and non-canonical pathways that mediate several aspects of brain development and function. Reelin's dimeric central fragment (CF), generated through proteolytic cleavage, is required for the lipoprotein-receptor-dependent canonical pathway activation. Here, we analyze the signaling properties of a variety of Reelin fragments and measure the differential binding affinities of monomeric and dimeric CF fragments to lipoprotein receptors to investigate the mode of canonical signal activation. We also present the cryoelectron tomography-solved dimeric structure of Reelin CF and support it using several other biophysical techniques. Our findings suggest that Reelin CF forms a covalent parallel dimer with some degree of flexibility between the two protein chains. As a result of this conformation, Reelin binds to lipoprotein receptors in a manner inaccessible to its monomeric form and is capable of stimulating canonical pathway signaling.

Keywords: AUC; ApoER2; Reelin; SAXS; VLDLR; cryo-ET; dimer; high-content analysis; structure-function.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cryoelectron Microscopy
HEK293 Cells
Humans
Protein Domains
Protein Multimerization
Receptors, LDL / metabolism
Reelin Protein / chemistry*
Reelin Protein / metabolism
Signal Transduction

Substances

Receptors, LDL
Reelin Protein
VLDL receptor
RELN protein, human

Grants and funding

P30 GM124166/GM/NIGMS NIH HHS/United States