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Cited by 9Connective tissue disorders researchProtein Structure and DynamicsCellular Mechanics and Interactions

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Microfibril-associated glycoprotein 4 forms octamers that mediate interactions with elastogenic proteins and cells

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Wozny, Michael R., Nelea, Valentin, Siddiqui, Iram Fatima S., Wanga, Shaynah, de Waard, Vivian, Strauss, Mike, Reinhardt, Dieter P.·2024·Nature Communications
DOI 10.1038/s41467-024-48377-zPMC PMC11091212

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Abstract

Microfibril-associated glycoprotein 4 (MFAP4) is a 36-kDa extracellular matrix glycoprotein with critical roles in organ fibrosis, chronic obstructive pulmonary disease, and cardiovascular disorders, including aortic aneurysms. MFAP4 multimerises and interacts with elastogenic proteins, including fibrillin-1 and tropoelastin, and with cells via integrins. Structural details of MFAP4 and its potential interfaces for these interactions are unknown. Here, we present a cryo-electron microscopy structure of human MFAP4. In the presence of calcium, MFAP4 assembles as an octamer, where two sets of homodimers constitute the top and bottom halves of each octamer. Each homodimer is linked together by an intermolecular disulphide bond. A C34S missense mutation prevents disulphide-bond formation between monomers but does not prevent octamer assembly. The atomic model, built into the 3.55 Å cryo-EM map, suggests that salt-bridge interactions mediate homodimer assembly, while non-polar residues form the interface between octamer halves. In the absence of calcium, an MFAP4 octamer dissociates into two tetramers. Binding studies with fibrillin-1, tropoelastin, LTBP4, and small fibulins show that MFAP4 has multiple surfaces for protein-protein interactions, most of which depend upon MFAP4 octamer assembly. The C34S mutation does not affect these protein interactions or cell interactions. MFAP4 assemblies with fibrillin-1 abrogate MFAP4 interactions with cells.

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Research works printed but not linked: 55

  1. Kobayashi, R. et al. Isolation and characterization of a new 36-kDa microfibril-associated glycoprotein from porcine aorta. J Biol Chem 264, 17437–17444 (1989). · DOI
  2. Wulf-Johansson, H. et al. Localization of microfibrillar-associated protein 4 (MFAP4) in human tissues: clinical evaluation of serum MFAP4 and its association with various cardiovascular conditions. PLoS One 8, e82243 (2013). · DOI
  3. Kanaan, R., Medlej-Hashim, M., Jounblat, R., Pilecki, B. & Sorensen, G. L. Microfibrillar-associated protein 4 in health and disease. Matrix Biol 111, 1–25 (2022). · DOI
  4. Sakai, L. Y., Keene, D. R. & Engvall, E. Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J. Cell Biol. 103, 2499–2509 (1986). · DOI
  5. Sabatier, L. et al. Fibrillin assembly requires fibronectin. Mol. Biol. Cell 20, 846–858 (2009). · DOI
  6. Kinsey, R. et al. Fibrillin-1 microfibril deposition is dependent on fibronectin assembly. J. Cell Sci. 121, 2696–2704 (2008). · DOI
  7. Hubmacher, D. et al. Biogenesis of extracellular microfibrils: multimerization of the fibrillin-1 C-terminus into bead-like structures enables self-assembly. Proc. Natl. Acad. Sci. USA 105, 6548–6553 (2008). · DOI
  8. Broekelmann, T. J. et al. Tropoelastin interacts with cell-surface glycosaminoglycans via its COOH-terminal domain. J. Biol. Chem. 280, 40939–40947 (2005). · DOI · Paper page
  9. Bax, D. V., Rodgers, U. R., Bilek, M. M. & Weiss, A. S. Cell adhesion to tropoelastin is mediated via the C-terminal GRKRK motif and integrin alphaVbeta3. J. Biol. Chem. 284, 28616–28623 (2009). · DOI
  10. Pilecki, B. et al. Characterization of microfibrillar-associated Protein 4 (MFAP4) as a tropoelastin- and fibrillin-binding protein involved in elastic fiber formation. J. Biol. Chem. 291, 1103–1114 (2016). · DOI
  11. Kumra, H. et al. Fibulin-4 exerts a dual role in LTBP-4L-mediated matrix assembly and function. Proc Natl Acad Sci USA 116, 20428–20437 (2019). · DOI
  12. Mecham, R. P. Elastin in lung development and disease pathogenesis. Matrix Biol 73, 6–20 (2018). · DOI · Paper page
  13. Papke, C. L. & Yanagisawa, H. Fibulin-4 and fibulin-5 in elastogenesis and beyond: insights from mouse and human studies. Matrix Biol. 37, 142–149 (2014). · DOI
  14. Sato, F. et al. Lysyl oxidase enhances the deposition of tropoelastin through the catalysis of tropoelastin molecules on the cell surface. Biol. Pharm. Bull. 40, 1646–1653 (2017). · DOI
  15. Godwin, A. R. F. et al. The role of fibrillin and microfibril binding proteins in elastin and elastic fibre assembly. Matrix Biol. 84, 17–30 (2019). · DOI
  16. Schiavinato, A. et al. Targeting of EMILIN-1 and EMILIN-2 to fibrillin microfibrils facilitates their incorporation into the extracellular matrix. J Invest Dermatol 136, 1150–1160 (2016). · DOI
  17. Ross, R. & Bornstein, P. The elastic fiber: the separation and partial characterization of its macromolecular components. J Cell Biol 40, 366–381 (1969). · DOI
  18. Toyoshima, T. et al. Ultrastructural distribution of 36-kD microfibril-associated glycoprotein (MAGP-36) in human and bovine tissues. J. Histochem. Cytochem. 47, 1049–1056 (1999). · DOI
  19. Toyoshima, T. et al. Differential gene expression of 36-kDa microfibril-associated glycoprotein (MAGP-36/MFAP4) in rat organs. Cell Tissue Res. 332, 271–278 (2008). · DOI
  20. Kasamatsu, S. et al. Essential role of microfibrillar-associated protein 4 in human cutaneous homeostasis and in its photoprotection. Sci. Rep. 1, 164 (2011). · DOI
  21. Holm, A. T. et al. Characterization of spontaneous air space enlargement in mice lacking microfibrillar-associated protein 4. Am. J. Physiol. Lung Cell Mol. Physiol. 308, L1114–L1124 (2015). · DOI
  22. Schlosser, A. et al. MFAP4 promotes vascular smooth muscle migration, proliferation and accelerates neointima formation. Arterioscler Thromb Vasc. Biol. 36, 122–133 (2016). · DOI
  23. Johansson, S. L. et al. Microfibrillar-associated protein 4: a potential biomarker of chronic obstructive pulmonary disease. Respir Med 108, 1336–1344 (2014). · DOI
  24. Kidholm, C. L. et al. Preliminary analysis of proteome alterations in non-aneurysmal, internal mammary artery tissue from patients with abdominal aortic aneurysms. PLoS One 13, e0192957 (2018). · DOI
  25. Bracht, T. et al. Analysis of disease-associated protein expression using quantitative proteomics-fibulin-5 is expressed in association with hepatic fibrosis. J. Proteome Res. 14, 2278–2286 (2015). · DOI
  26. Schlosser, A. et al. Microfibril-associated protein 4 binds to surfactant protein A (SP-A) and colocalizes with SP-A in the extracellular matrix of the lung. Scand. J. Immunol. 64, 104–116 (2006). · DOI
  27. Djokic, J., Fagotto-Kaufmann, C., Bartels, R., Nelea, V. & Reinhardt, D. P. Fibulin-3, -4, and -5 are highly susceptible to proteolysis, interact with cells and heparin, and form multimers. J. Biol. Chem. 288, 22821–22835 (2013). · DOI
  28. Osada, Y. et al. The fibrinogen C-terminal domain is seldom C-mannosylated but its C-mannosylation is important for the secretion of microfibril-associated glycoprotein 4. Biochim Biophys Acta - Gen Subj 1864, 129637 (2020). · DOI
  29. Toyoshima, T., Nishi, N., Kusama, H., Kobayashi, R. & Itano, T. 36-kDa microfibril-associated glycoprotein (MAGP-36) is an elastin-binding protein increased in chick aortae during development and growth. Exp. Cell Res. 307, 224–230 (2005). · DOI
  30. Shrive, A. K. et al. Crystal structure of the tetrameric fibrinogen-like recognition domain of fibrinogen C domain containing 1 (FIBCD1) protein. J Biol Chem 289, 2880–2887 (2014). · DOI
  31. Yu, X. et al. Structural basis for angiopoietin-1-mediated signaling initiation. Proc. Natl. Acad. Sci. USA 110, 7205–7210 (2013). · DOI
  32. Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta. Crystallogr. D Biol. Crystallogr. 66, 486–501 (2010). · DOI · Paper page
  33. Liebschner, D. et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta. Crystallogr. D Struct. Biol. 75, 861–877 (2019). · DOI · Paper page
  34. Kobayashi, R., Mizutani, A. & Hidaka, H. Isolation and characterization of a 36-kDa microfibril-associated glycoprotein by the newly synthesized isoquinolinesulfonamide affinity chromatography. Biochem Biophys Res Commun 198, 1262–1266 (1994). · DOI
  35. Yariv, B. et al. Using evolutionary data to make sense of macromolecules with a “face-lifted” ConSurf. Protein Sci 32, e4582 (2023). · DOI
  36. Sugar, S. et al. Alterations in protein expression and site-specific N-glycosylation of prostate cancer tissues. Sci Rep 11, 15886 (2021). · DOI
  37. Kucukelbir, A., Sigworth, F. J. & Tagare, H. D. Quantifying the local resolution of cryo-EM density maps. Nat Methods 11, 63–65 (2014). · DOI
  38. Dai, M., Dong, Z., Xu, K. & Zhang, Q. C. CryoRes: Local resolution estimation of cryo-EM density maps by deep learning. J. Mol. Biol. 435, 168059 (2023). · DOI
  39. Kimanius, D., Dong, L., Sharov, G., Nakane, T. & Scheres, S. H. W. New tools for automated cryo-EM single-particle analysis in RELION-4.0. Biochem. J. 478, 4169–4185 (2021). · DOI
  40. Nakane, T. & Scheres, S. H. W. Multi-body refinement of cryo-EM images in RELION. Methods Mol. Biol. 2215, 145–160 (2021). · DOI
  41. Garlatti, V. et al. Structural insights into the innate immune recognition specificities of L- and H-ficolins. EMBO J 26, 623–633 (2007). · DOI
  42. Canty, E. G. et al. Coalignment of plasma membrane channels and protrusions (fibripositors) specifies the parallelism of tendon. J Cell Biol. 165, 553–563 (2004). · DOI
  43. Zeyer, K. A., Zhang, R. M., Kumra, H., Hassan, A. & Reinhardt, D. P. The fibrillin-1 RGD integrin binding site regulates gene expression and cell function through microRNAs. J. Mol. Biol. 431, 401–421 (2019). · DOI
  44. Kaur, J. & Reinhardt, D. P. Immobilized metal affinity chromatography co-purifies TGF-beta1 with histidine-tagged recombinant extracellular proteins. PLoS One 7, e48629 (2012). · DOI
  45. Lin, G. et al. Homo- and heterotypic fibrillin-1 and -2 interactions constitute the basis for the assembly of microfibrils. J. Biol. Chem. 277, 50795–50804 (2002). · DOI
  46. Jensen, S. A., Reinhardt, D. P., Gibson, M. A. & Weiss, A. S. Protein interaction studies of MAGP-1 with tropoelastin and fibrillin-1. J. Biol. Chem. 276, 39661–39666 (2001). · DOI
  47. Zheng, S. Q. et al. MotionCor2: anisotropic correction of beam-induced motion for improved cryo-electron microscopy. Nat. Methods 14, 331–332 (2017). · DOI
  48. Rohou, A. & Grigorieff, N. CTFFIND4: fast and accurate defocus estimation from electron micrographs. J. Struct. Biol. 192, 216–221 (2015). · DOI
  49. Hohn, M. et al. SPARX, a new environment for Cryo-EM image processing. J. Struct. Biol. 157, 47–55 (2007). · DOI
  50. Xu, K., Wang, Z., Shi, J., Li, H. & Zhang, Q. C. A2-Net: molecular structure estimation from cryo-EM density volumes. 33rd. AAAI Conference on Artificial Intelligence 33, 1230–1237 (2019). · DOI
  51. Pettersen, E. F. et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021). · DOI
  52. Pettersen, E. F. et al. UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem 25, 1605–1612 (2004). · DOI
  53. Jiang, J. S. & Brünger, A. T. Protein hydration observed by X-ray diffraction. Solvation properties of penicillopepsin and neuraminidase crystal structures. J Mol Biol 243, 100–115 (1994). · DOI
  54. Humphries, M. J. Cell adhesion assays. Methods Mol Biol 522, 203–210 (2009). · DOI
  55. Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9, 671–675 (2012). · DOI · Paper page

Metadata

Authors
Wozny, M. R., Nelea, V., Siddiqui, I. F. S., Wanga, S., de Waard, V., Strauss, M., & Reinhardt, D. P.
Affiliations
  • McGill University (CA)
  • Amsterdam UMC Location University of Amsterdam (NL)
Corresponding author
  • Strauss, Mike
  • Reinhardt, Dieter P.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2024-05-13
Volume
15
Issue
1
Pages
4015-4015
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-024-48377-z
PMC
PMC11091212
OpenAlex
W4396871334
Cited by
9
References
2
Citations by year
2026: 3 · 2025: 6
FWCI
2.38
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • eScholarship@McGill (McGill) (submittedVersion, cc-by)
  • Pure Amsterdam UMC (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Gouvernement du Canada | Natural Sciences and Engineering Research Council of Canada (RGPIN-2022-05045, RGPIN-2020-04837)
  • Genetic Aortic Disorders Association Canada
  • Gouvernement du Canada | Canadian Institutes of Health Research (MFE-187851, PJT-186194)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Connective tissue disorders research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
Keywords
MFAP4, homodimer, disulfide bonds, salt bridges, protein-protein interaction, fibrillin-1, tropoelastin, LTBP4, fibulin, integrin binding, extracellular matrix glycoprotein, organ fibrosis, aortic aneurysm, chronic obstructive pulmonary disease
MeSH terms
Fibrillin-1, Calcium, Carrier Proteins, Glycoproteins, Humans, Models, Molecular, Protein Binding, Tropoelastin, Extracellular Matrix Proteins, Mutation, Missense, Cryoelectron Microscopy, Microfibrils, Adipokines, Protein Multimerization, HEK293 Cells

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