Structural Analysis of Cartilage Proteoglycans and Glycoproteins Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Zaia, Joseph; Liu, Beishan; Boynton, Raymond E.; Barry, Frank

doi:10.1006/abio.1999.4379

Cited by 18 publications

(17 citation statements)

References 55 publications

(46 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In Fig. 6b, oversulfated Z 19 and Y 19 ions having the structure of GalNAc [GlcA-GalNAc] 9, decorated with 11 SO 3 2 groups, were detected as triply charged species at m/z 1491.14 and 1497.13, respectively, accompanied by the corresponding sequences from the reducing end B 19 (9S) and C 19 (9S) showing both a regular sulfation distribution. This diagnostic set of ions along with the rest of regularly and oversulfated fragment ions identified in the complex spectrum from Fig.…”

Section: Resultsmentioning

confidence: 96%

On‐line sheathless capillary electrophoresis/nanoelectrospray ionization‐tandem mass spectrometry for the analysis of glycosaminoglycan oligosaccharides

Zamfir¹,

Seidler

Schönherr

et al. 2004

Electrophoresis

View full text Add to dashboard Cite

A novel approach in glycosaminoglycomics, based on sheathless on-line capillary electrophoresis/nanoelectrospray ionization-quadrupole time of flight-mass spectrometry (CE/nanoESI-QTOF-MS) and tandem MS of extended chondroitin sulfate/dermatan (CS/DS) oligosaccharide chains is described. The methodology required the construction of a new sheathless CE/nanoESI-QTOF-MS configuration, its implementation and optimization for the high sensitivity analysis of CS/DS oligosaccharide mixtures from conditioned culture medium of decorin transfected human embryonic kidney (HEK) 293 cells. Under newly established sheathless on-line CE/(-)nanoESI conditions for glycosaminoglycan (GAG) ionization and MS detection, single CS/DS oligosaccharide components of extended chain length and increased sulfation degree were identified. Molecular ions corresponding to species carrying 5 and 6 negative charges could be generated for large GAG oligosaccharide species in the negative ion nanoESI-MS. The optimized on-line conditions enabled the detection of molecular ions assigned to oversulfated tetradeca-, octadeca-, and eicosasaccharide CS/DS molecules, which represent the category of largest sulfated GAG-derived oligosaccharides evidenced by CE/ESI-MS. By on-line CE/ESI tandem MS in data-dependent acquisition mode the oversulfated eicosasaccharide species could be sequenced and the localization of the additional sulfate group along the chain could be determined.

show abstract

Section: Resultsmentioning

confidence: 96%

On‐line sheathless capillary electrophoresis/nanoelectrospray ionization‐tandem mass spectrometry for the analysis of glycosaminoglycan oligosaccharides

Zamfir¹,

Seidler

Schönherr

et al. 2004

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…An aliquot of the extracted peptide mixture was diluted 1:10 with ␣-cyano-4-hydroxycinnamic acid (CHCA, Hewlett-Packard) and used to acquire MALDI spectra as described (17).…”

Section: Methodsmentioning

confidence: 99%

The SH-3 and SH-4 Antibodies Recognize Distinct Epitopes on CD73 from Human Mesenchymal Stem Cells

Barry

Boynton

Murphy

et al. 2001

Biochemical and Biophysical Research Communications

Self Cite

223

102

View full text Add to dashboard Cite

“…The influence of hyperosmotic treatment on cell aggrecan protein expression, of interest based on our previous finding of increased chondrocyte aggrecan gene expression under similar treatments, could not be assessed here, since the sample preparation methods used were incompatible with processing insoluble polymers. 45 …”

Section: Discussionmentioning

confidence: 99%

Label-Free Protein Profiling of Adipose-Derived Human Stem Cells under Hyperosmotic Treatment

et al. 2011

View full text Add to dashboard Cite

Our previous work suggested that treatment of cells with hyperosmotic media during 2D passaging primes cells for cartilage tissue engineering applications. Here, we used label-free proteomic profiling to evaluate the effects of control and hyperosmotic treatment environments on the phenotype of multipotent adipose-derived stem cells (ASCs) cultivated with a chondrogenic growth factor cocktail. Spectra were recorded in a data-independent fashion at alternate low (precursor) and high (product) fragmentation voltages (MSE). This method was supplemented with data mining of accurate mass and retention time matches in precursor ion spectra across the experiment. The results indicated a complex cellular response to osmotic treatment, with a number of proteins differentially expressed between control and treated cell groups. The roles of some of these proteins have been documented in the literature as characteristic of the physiological states studied, especially aldose reductase (osmotic stress). This protein acted as a positive control in this work, providing independent corroborative validation. Other proteins, including 5’-nucleotidase and transgelin, have been previously linked to cell differentiation state. This study demonstrates that label-free profiling can serve as a useful tool in characterizing cellular responses to chondrogenic treatment regimes, recommending its use in optimization of cell priming protocols for cartilage tissue engineering.

show abstract

Structural Analysis of Cartilage Proteoglycans and Glycoproteins Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Cited by 18 publications

References 55 publications

On‐line sheathless capillary electrophoresis/nanoelectrospray ionization‐tandem mass spectrometry for the analysis of glycosaminoglycan oligosaccharides

On‐line sheathless capillary electrophoresis/nanoelectrospray ionization‐tandem mass spectrometry for the analysis of glycosaminoglycan oligosaccharides

The SH-3 and SH-4 Antibodies Recognize Distinct Epitopes on CD73 from Human Mesenchymal Stem Cells

Label-Free Protein Profiling of Adipose-Derived Human Stem Cells under Hyperosmotic Treatment

Contact Info

Product

Resources

About