Stable recombinant production of codon‐scrambled lubricin and mucin in human cells

Shurer, Carolyn R.; Wang, Yuyan; Feeney, Elizabeth Ann; Head, Shelby E.; Zhang, Victoria X.; C, Zhu; Stark, Morgan A.; Bonassar, Lawrence J.; Reesink, Heidi L.; Paszek, Matthew J.

doi:10.1002/bit.26940

Cited by 12 publications

(14 citation statements)

References 43 publications

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“…Purified rh-lubricin exhibited a molecular weight around 460 kDa (Figure S5A, Supporting Information), similar in size to endogenous lubricin and recombinant human lubricin generated by other laboratories. [16,38] Additionally, DLS measurement showed that the hydrodynamic diameter of purified rh-lubricin was 65.4 ± 1.1 nm (Figure S5C, Supporting Information), consistent with previous reports. [39] A lectin blot using FITC-Jacalin also confirmed the presence of O-glycans on purified rh-lubricin (Figure S5B, Supporting Information).…”

Section: Human Recombinant Lubricin Adsorption Restored Lubrication F...supporting

confidence: 90%

Mucin‐Like Glycoproteins Modulate Interfacial Properties of a Mimetic Ocular Epithelial Surface

et al. 2021

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Dry eye disease (DED) has high personal and societal costs, but its pathology remains elusive due to intertwined biophysical and biochemical processes at the ocular surface. Specifically, mucin deficiency is reported in a subset of DED patients, but its effects on ocular interfacial properties remain unclear. Herein a novel in vitro mucin‐deficient mimetic ocular surface (Mu‐DeMOS) with a controllable amount of membrane‐tethered mucin molecules is developed to represent the diseased ocular surfaces. Contact angle goniometry on mimetic ocular surfaces reveals that high surface roughness, but not the presence of hydrophilic mucin molecules, delivers constant hydration over native ocular surface epithelia. Live‐cell rheometry confirms that the presence of mucin‐like glycoproteins on ocular epithelial cells reduces shear adhesive strength at cellular interfaces. Together, optimal surface roughness and surface chemistry facilitate sustainable lubrication for healthy ocular surfaces, while an imbalance between them contributes to lubrication‐related dysfunction at diseased ocular epithelial surfaces. Furthermore, the restoration of low adhesive strength at Mu‐DeMOS interfaces through a mucin‐like glycoprotein, recombinant human lubricin, suggests that increased frictional damage at mucin‐deficient cellular surfaces may be reversible. More broadly, these results demonstrate that Mu‐DeMOS is a promising platform for drug screening assays and fundamental studies on ocular physiology.

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Section: Human Recombinant Lubricin Adsorption Restored Lubrication F...supporting

confidence: 90%

Mucin‐Like Glycoproteins Modulate Interfacial Properties of a Mimetic Ocular Epithelial Surface

et al. 2021

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“…Although several important papers have shown that the advantageous properties of mucuses can be emulated with synthetics, ,− the field of synthetic mucus is currently in its infancy. We hope that the efforts of CAMP and others already working to understand the structures and properties of mucuses ,− will support efforts to design biomimetic materials that seek to emulate the remarkable properties of these secretions found throughout the animal kingdom.…”

Section: Introductionmentioning

confidence: 98%

Comparative Animal Mucomics: Inspiration for Functional Materials from Ubiquitous and Understudied Biopolymers

Cerullo

Lai

Allam

et al. 2020

ACS Biomater. Sci. Eng.

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The functions of secreted animal mucuses are remarkably diverse and include lubricants, wet adhesives, protective barriers, and mineralizing agents. Although present in all animals, many open questions related to the hierarchical architectures, material properties, and genetics of mucus remain. Here, we summarize what is known about secreted mucus structure, describe the work of research groups throughout the world who are investigating various animal mucuses, and relate how these studies are revealing new mucus properties and the relationships between mucus hierarchical structure and hydrogel function. Finally, we call for a more systematic approach to studying animal mucuses so that data sets can be compared, omics-style, to address unanswered questions in the emerging field of mucomics. One major result that we anticipate from these efforts is design rules for creating new materials that are inspired by the structures and functions of animal mucuses.

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“…More recently, larger-scale biomanufacturing of human mucins utilized a codon-scrambling strategy to generate synonymous genes of two mucins of commercial interest in Freestyle 293-F cells. Methods for cDNA design and mucin production in mammalian host production systems were established (Shurer et al, 2019).…”

Section: Production Of Mucins and Mucin-like Molecules And Their Use mentioning

confidence: 99%

Mucins and Pathogenic Mucin-Like Molecules Are Immunomodulators During Infection and Targets for Diagnostics and Vaccines

2019

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Mucins and mucin-like molecules are highly O-glycosylated proteins present on the cell surface of mammals and other organisms. These glycoproteins are highly diverse in the apoprotein and glycan cores and play a central role in many biological processes and diseases. Mucins are the most abundant macromolecules in mucus and are responsible for its biochemical and biophysical properties. Mucin-like molecules cover various protozoan parasites, fungi and viruses. In humans, modifications in mucin glycosylation are associated with tumors in epithelial tissue. These modifications allow the distinction between normal and abnormal cell conditions and represent important targets for vaccine development against some cancers. Mucins and mucin-like molecules derived from pathogens are potential diagnostic markers and targets for therapeutic agents. In this review, we summarize the distribution, structure, role as immunomodulators, and the correlation of human mucins with diseases and perform a comparative analysis of mucins with mucin-like molecules present in human pathogens. Furthermore, we review the methods to produce pathogenic and human mucins using chemical synthesis and expression systems. Finally, we present applications of mucin-like molecules in diagnosis and prevention of relevant human diseases.

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Stable recombinant production of codon‐scrambled lubricin and mucin in human cells

Cited by 12 publications

References 43 publications

Mucin‐Like Glycoproteins Modulate Interfacial Properties of a Mimetic Ocular Epithelial Surface

Mucin‐Like Glycoproteins Modulate Interfacial Properties of a Mimetic Ocular Epithelial Surface

Comparative Animal Mucomics: Inspiration for Functional Materials from Ubiquitous and Understudied Biopolymers

Mucins and Pathogenic Mucin-Like Molecules Are Immunomodulators During Infection and Targets for Diagnostics and Vaccines

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