Effect of Disulfide Bonding and Multimerization on Proteoglycan 4's Cartilage Boundary Lubricating Ability and Adsorption

Overview

Journal Connect Tissue Res

Publisher Informa Healthcare

Date 2015 Dec 4

PMID 26631309

Citations 7

Authors

Saleem Abubacker

Dragana Ponjevic

Hyun O Ham

Phillip B Messersmith

John R Matyas

Tannin A Schmidt

Affiliations

Soon will be listed here.

Abstract

Purpose: The objectives of this study were to assess the cartilage boundary lubricating ability of (1) nonreduced (NR) disulfide-bonded proteoglycan 4 (PRG4) multimers versus PRG4 monomers and (2) NR versus reduced and alkylated (R/A) PRG4 monomers and to assess (3) the ability of NR PRG4 multimers versus monomers to adsorb to an articular cartilage surface.

Materials And Methods: PRG4 was separated into two preparations, PRG4 multimer enriched (PRG4Multi+) and PRG4 multimer deficient (PRG4Multi-), using size exclusion chromatography (SEC) and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The cartilage boundary lubricating ability of PRG4Multi+ and PRG4Multi- was compared at a physiological concentration (450 μg/mL) and assessed over a range of concentrations (45, 150, and 450 μg/mL). R/A and NR PRG4Multi- were evaluated at 450 μg/mL. Immunohistochemistry with anti-PRG4 antibody 4D6 was performed to visualize the adsorption of PRG4 preparations to the surface of articular cartilage explants.

Results: Separation into enriched populations of PRG4Multi+ and PRG4Multi- was achieved using SEC and was confirmed by SDS-PAGE. PRG4Multi+ and PRG4Multi- both functioned as effective friction-reducing cartilage boundary lubricants at 450 μg/mL, with PRG4Multi+ being more effective than PRG4Multi-. PRG4Multi+ lubricated in a dose-dependent manner, however, PRG4Multi- did not. R/A PRG4Multi- lubricated similar to NR PRG4Multi-. PRG4-containing solutions showed 4D6 immunoreactivity at the articular surface; the immunoreactive intensity of PRG4Multi+ appeared to be similar to SF, whereas PRG4Multi- appeared to have less intensity.

Conclusions: These results demonstrate that the intermolecular disulfide-bonded multimeric structure of PRG4 is important for its ability to adsorb to a cartilage surface and function as a boundary lubricant. These findings contribute to a greater understanding of the molecular basis of cartilage boundary lubrication of PRG4. Elucidating the PRG4 structure-lubrication function relationship will further contribute to the understanding of PRG4's role in diarthrodial joint homeostasis and disease.

Citing Articles

Genetic variation analysis of Guanling cattle based on whole-genome resequencing.

Xu L, Wang X, Wu J, Wang H, Zhou W, Liu J Anim Biosci. 2024; 37(12):2044-2053.

PMID: 38938024 PMC: 11541021. DOI: 10.5713/ab.24.0181.

O-glycans Expand Lubricin and Attenuate Its Viscosity and Shear Thinning.

Boushehri S, Holey H, Brosz M, Gumbsch P, Pastewka L, Aponte-Santamaria C Biomacromolecules. 2024; 25(7):3893-3908.

PMID: 38815979 PMC: 11238335. DOI: 10.1021/acs.biomac.3c01348.

A Mucin-Deficient Ocular Surface Mimetic Platform for Interrogating Drug Effects on Biolubrication, Antiadhesion Properties, and Barrier Functionality.

Madl A, Liu C, Cirera-Salinas D, Fuller G, Myung D ACS Appl Mater Interfaces. 2022; 14(16):18016-18030.

PMID: 35416028 PMC: 9052192. DOI: 10.1021/acsami.1c22280.

Proteomics Analysis of Tears and Saliva From Sjogren's Syndrome Patients.

Das N, Menon N, de Almeida L, Woods P, Heynen M, Jay G Front Pharmacol. 2021; 12:787193.

PMID: 34950038 PMC: 8689002. DOI: 10.3389/fphar.2021.787193.

Recombinant human PRG4 (rhPRG4) suppresses breast cancer cell invasion by inhibiting TGFβ-Hyaluronan-CD44 signalling pathway.

Sarkar A, Chanda A, Regmi S, Karve K, Deng L, Jay G PLoS One. 2019; 14(7):e0219697.

PMID: 31361756 PMC: 6667139. DOI: 10.1371/journal.pone.0219697.

References

Nugent-Derfus G, Chan A, Schumacher B, Sah R . PRG4 exchange between the articular cartilage surface and synovial fluid. J Orthop Res. 2007; 25(10):1269-76. DOI: 10.1002/jor.20431. View

Marcelino J, Carpten J, Suwairi W, Gutierrez O, Schwartz S, Robbins C . CACP, encoding a secreted proteoglycan, is mutated in camptodactyly-arthropathy-coxa vara-pericarditis syndrome. Nat Genet. 1999; 23(3):319-22. DOI: 10.1038/15496. View

Steele B, Alvarez-Veronesi M, Schmidt T . Molecular weight characterization of PRG4 proteins using multi-angle laser light scattering (MALLS). Osteoarthritis Cartilage. 2012; 21(3):498-504. DOI: 10.1016/j.joca.2012.12.002. View

Abubacker S, Ham H, Messersmith P, Schmidt T . Cartilage boundary lubricating ability of aldehyde modified proteoglycan 4 (PRG4-CHO). Osteoarthritis Cartilage. 2012; 21(1):186-9. PMC: 3538920. DOI: 10.1016/j.joca.2012.09.016. View

Ludwig T, McAllister J, Lun V, Wiley J, Schmidt T . Diminished cartilage-lubricating ability of human osteoarthritic synovial fluid deficient in proteoglycan 4: Restoration through proteoglycan 4 supplementation. Arthritis Rheum. 2012; 64(12):3963-71. DOI: 10.1002/art.34674. View

Antonacci J, Schmidt T, Serventi L, Cai M, Shu Y, Schumacher B . Effects of equine joint injury on boundary lubrication of articular cartilage by synovial fluid: role of hyaluronan. Arthritis Rheum. 2012; 64(9):2917-26. PMC: 3424370. DOI: 10.1002/art.34520. View

Kwiecinski J, Dorosz S, Ludwig T, Abubacker S, Cowman M, Schmidt T . The effect of molecular weight on hyaluronan's cartilage boundary lubricating ability--alone and in combination with proteoglycan 4. Osteoarthritis Cartilage. 2011; 19(11):1356-62. DOI: 10.1016/j.joca.2011.07.019. View

Chawla K, Ham H, Nguyen T, Messersmith P . Molecular resurfacing of cartilage with proteoglycan 4. Acta Biomater. 2010; 6(9):3388-94. PMC: 2908720. DOI: 10.1016/j.actbio.2010.03.025. View

Schmidt T, Plaas A, Sandy J . Disulfide-bonded multimers of proteoglycan 4 PRG4 are present in normal synovial fluids. Biochim Biophys Acta. 2009; 1790(5):375-84. DOI: 10.1016/j.bbagen.2009.03.016. View

10.

Elsaid K, Fleming B, Oksendahl H, Machan J, Fadale P, Hulstyn M . Decreased lubricin concentrations and markers of joint inflammation in the synovial fluid of patients with anterior cruciate ligament injury. Arthritis Rheum. 2008; 58(6):1707-15. PMC: 2789974. DOI: 10.1002/art.23495. View

11.

Zappone B, Greene G, Oroudjev E, Jay G, Israelachvili J . Molecular aspects of boundary lubrication by human lubricin: effect of disulfide bonds and enzymatic digestion. Langmuir. 2007; 24(4):1495-508. DOI: 10.1021/la702383n. View

12.

Jay G, Tantravahi U, Britt D, Barrach H, Cha C . Homology of lubricin and superficial zone protein (SZP): products of megakaryocyte stimulating factor (MSF) gene expression by human synovial fibroblasts and articular chondrocytes localized to chromosome 1q25. J Orthop Res. 2001; 19(4):677-87. DOI: 10.1016/S0736-0266(00)00040-1. View

13.

Schmidt T, Gastelum N, Han E, Nugent-Derfus G, Schumacher B, Sah R . Differential regulation of proteoglycan 4 metabolism in cartilage by IL-1alpha, IGF-I, and TGF-beta1. Osteoarthritis Cartilage. 2007; 16(1):90-7. DOI: 10.1016/j.joca.2007.05.009. View

14.

Ikegawa S, Sano M, Koshizuka Y, Nakamura Y . Isolation, characterization and mapping of the mouse and human PRG4 (proteoglycan 4) genes. Cytogenet Cell Genet. 2000; 90(3-4):291-7. DOI: 10.1159/000056791. View

15.

Jay G, Torres J, Warman M, Laderer M, Breuer K . The role of lubricin in the mechanical behavior of synovial fluid. Proc Natl Acad Sci U S A. 2007; 104(15):6194-9. PMC: 1851076. DOI: 10.1073/pnas.0608558104. View

16.

Schmidt T, Gastelum N, Nguyen Q, Schumacher B, Sah R . Boundary lubrication of articular cartilage: role of synovial fluid constituents. Arthritis Rheum. 2007; 56(3):882-91. DOI: 10.1002/art.22446. View

17.

Perez-Vilar J, Mabolo R . Gel-forming mucins. Notions from in vitro studies. Histol Histopathol. 2007; 22(4):455-64. DOI: 10.14670/HH-22.455. View

18.

Jones A, Gleghorn J, Hughes C, Fitz L, Zollner R, Wainwright S . Binding and localization of recombinant lubricin to articular cartilage surfaces. J Orthop Res. 2006; 25(3):283-92. DOI: 10.1002/jor.20325. View

19.

Schmidt T, Sah R . Effect of synovial fluid on boundary lubrication of articular cartilage. Osteoarthritis Cartilage. 2006; 15(1):35-47. DOI: 10.1016/j.joca.2006.06.005. View

20.

Rhee D, Marcelino J, Al-Mayouf S, Schelling D, Bartels C, Cui Y . Consequences of disease-causing mutations on lubricin protein synthesis, secretion, and post-translational processing. J Biol Chem. 2005; 280(35):31325-32. DOI: 10.1074/jbc.M505401200. View