Proteomic identification of galectin-3 binding ligands and characterization of galectin-3 proteolytic cleavage in human prostasomes

Kovak, Matthew R.; Saraswati, Sarika; Goddard, S.; Diekman, Alan B.

doi:10.1111/j.2047-2927.2013.00099.x

Cited by 19 publications

(22 citation statements)

References 40 publications

(73 reference statements)

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“…Finally, affinity capture chromatography, coupled with LC‐MS/MS, identified KLK3 to interact with galectin‐3 on the surface of prostate‐expressed vesicles, or “prostasomes,” in seminal fluid. Galectin‐3 was confirmed as a novel KLK3 proteolytic substrate and proteolysis of this glycoprotein in seminal fluid was reduced following proteolytic inhibition of KLK3, hence galectin‐3 appears to be a definitive biological KLK3 substrate . This study constitutes another example where identification of a KLK substrate was determined in a reverse manner by a global screening approach used to identify binding proteins of what was eventually found to be a KLK substrate.…”

Section: Proteomics As a Tool For Global Analysis Of Klk Production Amentioning

confidence: 90%

Proteomic and other analyses to determine the functional consequences of deregulated kallikrein‐related peptidase (KLK) expression in prostate and ovarian cancer

Fuhrman-Luck

Silva

Dong

et al. 2014

Proteomics Clinical Apps

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Rapidly developing proteomic tools are improving detection of deregulated kallikrein-related peptidase (KLK) expression, at the protein level, in prostate and ovarian cancer, as well as facilitating the determination of functional consequences downstream. MS-driven proteomics uniquely allows for the detection, identification, and quantification of thousands of proteins in a complex protein pool, and this has served to identify certain KLKs as biomarkers for these diseases. In this review, we describe applications of this technology in KLK biomarker discovery and elucidate MS-based techniques that have been used for unbiased, global screening of KLK substrates within complex protein pools. Although MS-based KLK degradomic studies are limited to date, they helped to discover an array of novel KLK substrates. Substrates identified by MS-based degradomics are reported with improved confidence over those determined by incubating a purified or recombinant substrate and protease of interest, in vitro. We propose that these novel proteomic approaches represent the way forward for KLK research, in order to correlate proteolysis of biological substrates with tissue-related consequences, toward clinical targeting of KLK expression and function for cancer diagnosis, prognosis, and therapies.

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Section: Proteomics As a Tool For Global Analysis Of Klk Production Amentioning

confidence: 90%

Proteomic and other analyses to determine the functional consequences of deregulated kallikrein‐related peptidase (KLK) expression in prostate and ovarian cancer

Fuhrman-Luck

Silva

Dong

et al. 2014

Proteomics Clinical Apps

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show abstract

“…The subsequent supernatant was ultracentrifuged at 100,000 g at 4°C for 2 hr to remove membrane and particulate material. Recombinant human galectin‐3 was covalently bound to cyanogen bromide‐activated Sepharose (Sigma, St. Louis, MO, USA) following the manufacturer's instructions, and galectin‐3 affinity column chromatography was performed as previously described …”

Section: Methodsmentioning

confidence: 99%

“…Prostasomes are exosome‐like vesicles in seminal plasma that are secreted by the prostatic epithelium and are proposed to have immunomodulatory functions in the female reproductive tract . We characterized galectin‐3 binding ligands in human prostasomes and demonstrated that galectin‐3 is a proteolytic substrate for prostate‐specific antigen (PSA) in seminal plasma and on the surface of prostasomes . We now report the results of a proteomic study that investigated galectin‐3 function in seminal plasma by characterizing proteins that bound to immobilized galectin‐3.…”

Section: Introductionmentioning

confidence: 99%

Investigation of Galectin‐3 Function in the Reproductive Tract by Identification of Binding Ligands in Human Seminal Plasma

Kovak

Saraswati

Schoen

et al. 2014

American J Rep Immunol

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Problem Galectin-3 is a β-galactoside binding protein with immunomodulatory properties and exerts its extracellular functions via interactions with glycoconjugate ligands. Therefore, to elucidate the function of galectin-3, binding ligands in human seminal plasma were investigated. Method of Study Galectin-3 binding proteins were isolated from seminal plasma by affinity chromatography, and candidate ligands were identified by MS/MS. Biochemical methods were used to characterize the ability of galectin-3 to bind its ligands. Results Identified galectin-3 ligands included CD13, MUC6, PAP, PSA and ZAG. 1D and 2D electrophoretic analysis of seminal plasma demonstrated that CD13, PAP, PSA, and ZAG immunoreactivity co-migrated with galectin-3-reactive protein bands and spots at expected molecular weights and pIs. Inhibition assays indicated that CD13, PSA, PAP, and ZAG, interact with galectin-3 in a protein-carbohydrate manner. Conclusion The galectin-3 binding ligands identified in this study indicate multiple roles for galectin-3 in the reproductive and immunological functions of seminal plasma.

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“…This suggested that CLU binds specifically to disrupted areas of the barrier. CLU was identified as an interacting candidate in a recent proteomics screen for potential LGALS3 interacting proteins from human prostasomes [179].…”

Section: Ocular Surface Barrier Function In Dry Eye Diseasementioning

confidence: 99%

Clusterin in the eye: An old dog with new tricks at the ocular surface

Fini

Bauskar

Jeong

et al. 2016

Experimental Eye Research

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, M. R. (2016). Clusterin in the eye: an old dog with new tricks at the ocular surface. Experimental Eye Research, Clusterin in the eye: an old dog with new tricks at the ocular surface AbstractThe multifunctional protein clusterin (CLU) was first described in 1983 as a secreted glycoprotein present in ram rete testis fluid that enhanced aggregation ('clustering') of a variety of cells in vitro. It was also independently discovered in a number of other systems. By the early 1990s, CLU was known under many names and its expression had been demonstrated throughout the body, including in the eye. Its homeostatic activities in proteostasis, cytoprotection, and anti-inflammation have been well documented, however its roles in health and disease are still not well understood. CLU is prominent at fluid-tissue interfaces, and in 1996 it was demonstrated to be the most highly expressed transcript in the human cornea, the protein product being localized to the apical layers of the mucosal epithelia of the cornea and conjunctiva. CLU protein is also present in human tears. Using a preclinical mouse model for desiccating stress that mimics human dry eye disease, the authors recently demonstrated that CLU prevents and ameliorates ocular surface barrier disruption by a remarkable sealing mechanism dependent on attainment of a critical all-or-none concentration in the tears. When the CLU level drops below the critical all-or-none threshold, the barrier becomes vulnerable to desiccating stress. CLU binds selectively to the ocular surface subjected to desiccating stress in vivo, and in vitro to LGALS3 (galectin-3), a key barrier component. Positioned in this way, CLU not only physically seals the ocular surface barrier, but it also protects the barrier cells and prevents further damage to barrier structure. CLU depletion from the ocular surface epithelia is seen in a variety of inflammatory conditions in humans and mice that lead to squamous metaplasia and a keratinized epithelium. This suggests that CLU might have a specific role in maintaining mucosal epithelial differentiation, an idea that can now be tested using the mouse model for desiccating stress. Most excitingly, the new findings suggest that CLU could serve as a novel biotherapeutic for dry eye disease. Competing Interests: US patent 9,241,974 B2 entitled "Clusterin pharmaceuticals and treatment methods using the same" (inventors: MEF and SJ), assigned to the University of Southern California, is connected with this work. MEF holds a management position with Proteris Biotech, Inc., Pasadena, CA, which is developing Protearin for dry eye based on clusterin. MRW declares that he has no competing interests. Page 3 of 65 AbstractThe multifunctional protein clusterin (CLU) was first described in 1983 as a secreted glycoprotein present in ram rete testis fluid that enhanced aggregation ('clustering') of a variety of cells in vitro. It was also independently discovered in a number of other systems. By the early 1990s, CLU was known under many names and its expression had bee...

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Proteomic identification of galectin-3 binding ligands and characterization of galectin-3 proteolytic cleavage in human prostasomes

Cited by 19 publications

References 40 publications

Proteomic and other analyses to determine the functional consequences of deregulated kallikrein‐related peptidase (KLK) expression in prostate and ovarian cancer

Proteomic and other analyses to determine the functional consequences of deregulated kallikrein‐related peptidase (KLK) expression in prostate and ovarian cancer

Investigation of Galectin‐3 Function in the Reproductive Tract by Identification of Binding Ligands in Human Seminal Plasma

Clusterin in the eye: An old dog with new tricks at the ocular surface

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