Detection and identification of plasma proteins that bind GlialCAM using ProteinChip™ Arrays, SELDI‐TOF MS, and nano‐LC MS/MS

Favre-Kontula, Linda; Sattonnet-Roche, Pascale; Magnenat, Edith; Proudfoot, Amanda E. I.; Boschert, Ursula; Xenarios, Ioannis; Vilbois, Francis; Antonsson, Bruno

doi:10.1002/pmic.200700564

Cited by 16 publications

(15 citation statements)

References 33 publications

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“…HepaCAM was recently demonstrated to be differentially expressed in human hepatocellular carcinoma [13,14]. Studies have shown that the expression of hepaCAM is frequently lost in human hepatocellular carcinoma and processed a tumor suppressor gene [7,15,16].…”

Section: Discussionmentioning

confidence: 99%

Functional significance of the hepaCAM gene in bladder cancer

He¹,

Wu²,

Luo

et al. 2010

BMC Cancer

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BackgroundThe hepaCAM gene encodes a new immunoglobulin-like cell adhesion molecule, and its expression is suppressed in a variety of human cancers. Additionally, hepaCAM possesses properties often observed in tumor suppressor genes. However, the expression and biological function of hepaCAM has not been investigated in bladder cancer. Therefore we sought to examine hepaCAM expression and the relationship between its structure and function in human transitional cell carcinoma of bladder (TCCB).Materials and methodsHepaCAM expression was evaluated in 28 normal and 34 TCCB bladder specimens and 2 TCCB cell lines using semi-quantitative RT-PCR. The wild-type hepaCAM and the extracellular domain-truncated mutant gene were transfected into the TCCB cell line T24, and the biological properties of both the wild-type gene and the domain-truncated mutant were then assessed.ResultsHepaCAM expression was down-regulated in 82% (28/34) of TCCB specimens and undetectable in the 2 TCCB cell lines tested. The localization of hepaCAM appeared to be dependent on cell density in T24 cells. In widely spread cells, hepaCAM accumulated on the perinuclear membrane and the cell surface protrusions, whereas in confluent cells, hepaCAM was predominantly localized at the sites of cell-cell contacts on the cell membrane. Functionally, hepaCAM expressed not only increased cell spreading, delayed cell detachment, enhanced wound healing and increased cell invasion; it also inhibited cell growth (P < 0.01). When the extracellular domain was deleted, the localization of hepaCAM was significantly altered, and it lost both its adhesive function and its influence on cell growth.ConclusionsHepaCAM is involved in cell adhesion and growth control, and its expression is frequently silenced in TCCB. The extracellular domain of hepaCAM is essential to its physiological and biological functions.

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Section: Discussionmentioning

confidence: 99%

Functional significance of the hepaCAM gene in bladder cancer

He¹,

Wu²,

Luo

et al. 2010

BMC Cancer

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“…Initially analysis of the integrity of CCL18 on the NP20 ProteinChip was first determined by applying CCL18 (5 μl at a concentration of 1 mg/ml) onto a NP20 ProteinChip Array (BioRad) as previously described (24). A saturated solution of sinapinic acid in 50% acetonitrile containing 1% Trifluoroacetic acid (TFA) in distilled H 2 O was added onto each spot and air-dried.…”

Section: Methodsmentioning

confidence: 99%

The Activity of CCL18 is Principally Mediated through Interaction with Glycosaminoglycans

et al. 2013

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The CC chemokine ligand 18 (CCL18) was first identified as a chemoattractant for naïve T cells. It has been reported to recruit T and B lymphocytes, and we show here, natural killer (NK) cells, but with low efficacy. Investigation of its ability to elicit G-protein-coupled signaling showed that it does not involve extracellular signal-regulated kinase (ERK) phosphorylation, and it is not able to induce receptor internalization, as assessed on CCR3. CCL18 has recently been reported to possess activities unrelated to cellular recruitment, but it had no effect on T lymphocyte proliferation. We postulated that a more potent chemoattractant may be produced under inflammatory conditions but only minor truncations were observed, with the major form being the full-length protein. In view of the lack of potent immunomodulatory properties, we wondered if binding to CCL18 by the tick chemokine binding proteins Evasin-1 and -4 was an artifact of the methods used, but complex formation was confirmed by size exclusion chromatography, and abrogation of its binding to, and antagonism of, CCR3. Its receptor has remained elusive since its cloning in 1997, although it has been reported to induce migration of breast cancer cells by signaling through PITPNM3, but we show that this receptor is not expressed on lymphocytes. We have developed a radiolabeled equilibrium competition binding assay and demonstrated that it bound with high affinity to peripheral blood leukocytes (PBLs), but the binding was displaced similarly by both unlabelled CCL18 as well as heparin. Both heparin binding and binding to PBLs are considerably abrogated by mutation of the BBXB motif in the 40s loop suggesting an essential role of the CCL18-glycosaminoglycan interaction.

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“…The affinity reagent may be a protein, antibody, peptide, aptamer or another biomolecule (Tuerk & MacDougal-Waugh, 1993;Durham & Regnier, 2006;Cole et al, 2007;Csanky et al, 2007;Sennels et al, 2007;Dubois et al, 2008;Evans-Nguyen et al, 2008;Favre-Kontula et al, 2008). Immuno-affinity chromatography of specific antibodies immobilized to hydrazide resin and monitored by mass spectrometry has been shown to be in the ng/mL range and shows agreement with ELISA (Nicol et al, 2008) and may be sensitive to femtomol amounts on column (Keshishian et al, 2007;Kulasingam et al, 2008).…”

Section: Affinity Chromatographymentioning

confidence: 95%

Mass spectrometry of peptides and proteins from human blood

Zhu

Bowden

Zhang

et al. 2010

Mass Spectrometry Reviews

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It is difficult to convey the accelerating rate and growing importance of mass spectrometry applications to human blood proteins and peptides. Mass spectrometry can rapidly detect and identify the ionizable peptides from the proteins in a simple mixture and reveal many of their post-translational modifications. However, blood is a complex mixture that may contain many proteins first expressed in cells and tissues. The complete analysis of blood proteins is a daunting task that will rely on a wide range of disciplines from physics, chemistry, biochemistry, genetics, electromagnetic instrumentation, mathematics and computation. Therefore the comprehensive discovery and analysis of blood proteins will rank among the great technical challenges and require the cumulative sum of many of mankind's scientific achievements together. A variety of methods have been used to fractionate, analyze and identify proteins from blood, each yielding a small piece of the whole and throwing the great size of the task into sharp relief. The approaches attempted to date clearly indicate that enumerating the proteins and peptides of blood can be accomplished. There is no doubt that the mass spectrometry of blood will be crucial to the discovery and analysis of proteins, enzyme activities, and post-translational processes that underlay the mechanisms of disease. At present both discovery and quantification of proteins from blood are commonly reaching sensitivities of ∼1 ng/mL.

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Detection and identification of plasma proteins that bind GlialCAM using ProteinChip™ Arrays, SELDI‐TOF MS, and nano‐LC MS/MS

Cited by 16 publications

References 33 publications

Functional significance of the hepaCAM gene in bladder cancer

Functional significance of the hepaCAM gene in bladder cancer

The Activity of CCL18 is Principally Mediated through Interaction with Glycosaminoglycans

Mass spectrometry of peptides and proteins from human blood

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