Hyperphosphorylation of Human Osteopontin and Its Impact on Structural Dynamics and Molecular Recognition

Overview

Journal Biochemistry

Specialty Biochemistry

Date 2021 Apr 20

PMID 33876640

Citations 9

Authors

Borja Mateos

Julian Holzinger

Clara Conrad-Billroth

Gerald Platzer

Szymon Zerko

Marco Sealey-Cardona

Dorothea Anrather

Wiktor Kozminski

Robert Konrat

Affiliations

Soon will be listed here.

Abstract

Protein phosphorylation is an abundant post-translational modification (PTM) and an essential modulator of protein functionality in living cells. Intrinsically disordered proteins (IDPs) are particular targets of PTM protein kinases due to their involvement in fundamental protein interaction networks. Despite their dynamic nature, IDPs are far from having random-coil conformations but exhibit significant structural heterogeneity. Changes in the molecular environment, most prominently in the form of PTM via phosphorylation, can modulate these structural features. Therefore, how phosphorylation events can alter conformational ensembles of IDPs and their interactions with binding partners is of great interest. Here we study the effects of hyperphosphorylation on the IDP osteopontin (OPN), an extracellular target of the Fam20C kinase. We report a full characterization of the phosphorylation sites of OPN using a combined nuclear magnetic resonance/mass spectrometry approach and provide evidence for an increase in the local flexibility of highly phosphorylated regions and the ensuing overall structural elongation. Our study emphasizes the simultaneous importance of electrostatic and hydrophobic interactions in the formation of compact substates in IDPs and their relevance for molecular recognition events.

Citing Articles

Multi-Omics Profiling Identifies Microglial Annexin A2 as a Key Mediator of NF-κB Pro-inflammatory Signaling in Ischemic Reperfusion Injury.

Tian X, Yang W, Jiang W, Zhang Z, Liu J, Tu H Mol Cell Proteomics. 2024; 23(2):100723.

PMID: 38253182 PMC: 10879806. DOI: 10.1016/j.mcpro.2024.100723.

The Multifaceted Role of Osteopontin in Prostate Pathologies.

Silver S, Popovics P Biomedicines. 2023; 11(11).

PMID: 38001899 PMC: 10669591. DOI: 10.3390/biomedicines11112895.

The molecular basis for cellular function of intrinsically disordered protein regions.

Holehouse A, Kragelund B Nat Rev Mol Cell Biol. 2023; 25(3):187-211.

PMID: 37957331 PMC: 11459374. DOI: 10.1038/s41580-023-00673-0.

Intrinsically disordered regions are poised to act as sensors of cellular chemistry.

Moses D, Ginell G, Holehouse A, Sukenik S Trends Biochem Sci. 2023; 48(12):1019-1034.

PMID: 37657994 PMC: 10840941. DOI: 10.1016/j.tibs.2023.08.001.

Thrombin Cleavage of Osteopontin and the Host Anti-Tumor Immune Response.

Leung L, Myles T, Morser J Cancers (Basel). 2023; 15(13).

PMID: 37444590 PMC: 10340489. DOI: 10.3390/cancers15133480.

References

Ulrich E, Akutsu H, Doreleijers J, Harano Y, Ioannidis Y, Lin J . BioMagResBank. Nucleic Acids Res. 2007; 36(Database issue):D402-8. PMC: 2238925. DOI: 10.1093/nar/gkm957. View

Christensen B, Nielsen M, Haselmann K, Petersen T, Sorensen E . Post-translationally modified residues of native human osteopontin are located in clusters: identification of 36 phosphorylation and five O-glycosylation sites and their biological implications. Biochem J. 2005; 390(Pt 1):285-92. PMC: 1184582. DOI: 10.1042/BJ20050341. View

Mateos B, Sealey-Cardona M, Balazs K, Konrat J, Staffler G, Konrat R . NMR Characterization of Surface Receptor Protein Interactions in Live Cells Using Methylcellulose Hydrogels. Angew Chem Int Ed Engl. 2019; 59(10):3886-3890. PMC: 7065066. DOI: 10.1002/anie.201913585. View

Tian X, Azpurua J, Hine C, Vaidya A, Myakishev-Rempel M, Ablaeva J . High-molecular-mass hyaluronan mediates the cancer resistance of the naked mole rat. Nature. 2013; 499(7458):346-9. PMC: 3720720. DOI: 10.1038/nature12234. View

Raine J, Winter R, Davey A, Tucker S . Unknown syndrome: microcephaly, hypoplastic nose, exophthalmos, gum hyperplasia, cleft palate, low set ears, and osteosclerosis. J Med Genet. 1989; 26(12):786-8. PMC: 1015765. DOI: 10.1136/jmg.26.12.786. View

Takeda M, Terasawa H, Sakakura M, Yamaguchi Y, Kajiwara M, Kawashima H . Hyaluronan recognition mode of CD44 revealed by cross-saturation and chemical shift perturbation experiments. J Biol Chem. 2003; 278(44):43550-5. DOI: 10.1074/jbc.M308199200. View

Gibbs E, Lu F, Portz B, Fisher M, Medellin B, Laremore T . Phosphorylation induces sequence-specific conformational switches in the RNA polymerase II C-terminal domain. Nat Commun. 2017; 8:15233. PMC: 5437310. DOI: 10.1038/ncomms15233. View

Jakob U, Kriwacki R, Uversky V . Conditionally and transiently disordered proteins: awakening cryptic disorder to regulate protein function. Chem Rev. 2014; 114(13):6779-805. PMC: 4090257. DOI: 10.1021/cr400459c. View

Mittag T, Orlicky S, Choy W, Tang X, Lin H, Sicheri F . Dynamic equilibrium engagement of a polyvalent ligand with a single-site receptor. Proc Natl Acad Sci U S A. 2008; 105(46):17772-7. PMC: 2582940. DOI: 10.1073/pnas.0809222105. View

10.

Sodek J, Ganss B, McKee M . Osteopontin. Crit Rev Oral Biol Med. 2000; 11(3):279-303. DOI: 10.1177/10454411000110030101. View

11.

Bielska A, Zondlo N . Hyperphosphorylation of tau induces local polyproline II helix. Biochemistry. 2006; 45(17):5527-37. DOI: 10.1021/bi052662c. View

12.

Salih E, Ashkar S, Gerstenfeld L, Glimcher M . Identification of the phosphorylated sites of metabolically 32P-labeled osteopontin from cultured chicken osteoblasts. J Biol Chem. 1997; 272(21):13966-73. DOI: 10.1074/jbc.272.21.13966. View

13.

Theillet F, Smet-Nocca C, Liokatis S, Thongwichian R, Kosten J, Yoon M . Cell signaling, post-translational protein modifications and NMR spectroscopy. J Biomol NMR. 2012; 54(3):217-36. PMC: 4939263. DOI: 10.1007/s10858-012-9674-x. View

14.

Ishikawa H, Xu A, Ogura E, Manning G, Irvine K . The Raine syndrome protein FAM20C is a Golgi kinase that phosphorylates bio-mineralization proteins. PLoS One. 2012; 7(8):e42988. PMC: 3416761. DOI: 10.1371/journal.pone.0042988. View

15.

Antz C, Bauer T, Kalbacher H, Frank R, Covarrubias M, Kalbitzer H . Control of K+ channel gating by protein phosphorylation: structural switches of the inactivation gate. Nat Struct Biol. 1999; 6(2):146-50. DOI: 10.1038/5833. View

16.

Boskey A, Christensen B, Taleb H, Sorensen E . Post-translational modification of osteopontin: effects on in vitro hydroxyapatite formation and growth. Biochem Biophys Res Commun. 2012; 419(2):333-8. PMC: 3299831. DOI: 10.1016/j.bbrc.2012.02.024. View

17.

Hunter G, Kyle C, Goldberg H . Modulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin-mediated inhibition of hydroxyapatite formation. Biochem J. 1994; 300 ( Pt 3):723-8. PMC: 1138226. DOI: 10.1042/bj3000723. View

18.

Parigi G, Rezaei-Ghaleh N, Giachetti A, Becker S, Fernandez C, Blackledge M . Long-range correlated dynamics in intrinsically disordered proteins. J Am Chem Soc. 2014; 136(46):16201-9. DOI: 10.1021/ja506820r. View

19.

Martin E, Holehouse A, Grace C, Hughes A, Pappu R, Mittag T . Sequence Determinants of the Conformational Properties of an Intrinsically Disordered Protein Prior to and upon Multisite Phosphorylation. J Am Chem Soc. 2016; 138(47):15323-15335. PMC: 5675102. DOI: 10.1021/jacs.6b10272. View

20.

Kubota T, Zhang Q, Wrana J, Ber R, Aubin J, Butler W . Multiple forms of SppI (secreted phosphoprotein, osteopontin) synthesized by normal and transformed rat bone cell populations: regulation by TGF-beta. Biochem Biophys Res Commun. 1989; 162(3):1453-9. DOI: 10.1016/0006-291x(89)90837-1. View