scholarly journals Nanowell-mediated two-dimensional liquid chromatography enables deep proteome profiling of <1000 mammalian cells

2018 ◽  
Vol 9 (34) ◽  
pp. 6944-6951 ◽  
Author(s):  
Maowei Dou ◽  
Ying Zhu ◽  
Andrey Liyu ◽  
Yiran Liang ◽  
Jing Chen ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Mammalian Cells ◽  
Two Dimensional ◽  
Proteome Profiling ◽  
Peptide Separations

Miniaturized multidimensional peptide separations can greatly increase the coverage in proteome profiling for trace samples.

Application of on-line two-dimensional liquid chromatography in comparative proteome analysis of antlers with different growing stages

2010 ◽  
Vol 28 (2) ◽  
pp. 146-151 ◽  
Author(s):  
Liang GAO ◽  
Xiaoqiang QIAO ◽  
Zhen LIANG ◽  
Lihua ZHANG ◽  
Yushu HUO ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
On Line

scholarly journals Peak-tracking algorithm for use in comprehensive two-dimensional liquid chromatography – Application to monoclonal-antibody peptides

2021 ◽  
Vol 1639 ◽  
pp. 461922
Author(s):  
Stef R.A. Molenaar ◽  
Tina A. Dahlseid ◽  
Gabriel M. Leme ◽  
Dwight R. Stoll ◽  
Peter J. Schoenmakers ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Liquid Chromatography ◽  
Tracking Algorithm ◽  
Two Dimensional ◽  
Peak Tracking

Phospholipase D1b and D2a generate structurally identical phosphatidic acid species in mammalian cells

2001 ◽  
Vol 360 (3) ◽  
pp. 707-715 ◽  
Author(s):  
Trevor R. PETTITT ◽  
Mark McDERMOTT ◽  
Khalid M. SAQIB ◽  
Neil SHIMWELL ◽  
Michael J. O. WAKELAM
Keyword(s):  
Liquid Chromatography ◽  
Phosphatidic Acid ◽  
Phospholipase D ◽  
Mammalian Cells ◽  
Inducible Promoter ◽  
In Vitro Assays ◽  
Protein Levels ◽  
Intracellular Messenger

Mammalian cells contain different phospholipase D enzymes (PLDs) whose distinct physiological roles are poorly understood and whose products have not been characterized. The development of porcine aortic endothelial (PAE) cell lines able to overexpress PLD-1b or −2a under the control of an inducible promoter has enabled us to characterize both the substrate specificity and the phosphatidic acid (PtdOH) product of these enzymes under controlled conditions. Liquid chromatography–MS analysis showed that PLD1b- and PLD2a-transfected PAE cells, as well as COS7 and Rat1 cells, generate similar PtdOH and, in the presence of butan-1-ol, phosphatidylbutanol (PtdBut) profiles, enriched in mono- and di-unsaturated species, in particular 16:0/18:1. Although PtdBut mass increased, the species profile did not change in cells stimulated with ATP or PMA. Overexpression of PLD made little difference to basal or stimulated PtdBut formation, indicating that activity is tightly regulated in vivo and that factors other than just PLD protein levels limit hydrolytic function. In vitro assays using PLD-enriched lysates showed that the enzyme could utilize both phosphatidylcholine and, much less efficiently, phosphatidylethanolamine, with slight selectivity towards mono- and di-unsaturated species. Phosphatidylinositol was not a substrate. Thus PLD1b and PLD2a hydrolyse a structurally similar substrate pool to generate an identical PtdOH product enriched in mono- and di-unsaturated species that we propose to function as the intracellular messenger forms of this lipid.

Optimization strategies for off-line two-dimensional liquid chromatography

2009 ◽  
Vol 1216 (12) ◽  
pp. 2511-2518 ◽  
Author(s):  
Krisztián Horváth ◽  
Jacob Fairchild ◽  
Georges Guiochon
Keyword(s):  
Liquid Chromatography ◽  
Two Dimensional

scholarly journals Analyses of Lipid A Diversity in Gram-Negative Intestinal Bacteria Using Liquid Chromatography–Quadrupole Time-of-Flight Mass Spectrometry

Metabolites ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 197
Author(s):  
Nobuyuki Okahashi ◽  
Masahiro Ueda ◽  
Fumio Matsuda ◽  
Makoto Arita
Keyword(s):  
Mass Spectrometry ◽  
Immune Response ◽  
Liquid Chromatography ◽  
Mammalian Cells ◽  
Lipid A ◽  
Acyl Chain ◽  
Time Of Flight ◽  
Intestinal Bacteria ◽  
Gram Negative ◽  
Flight Mass Spectrometry

Lipid A is a characteristic molecule of Gram-negative bacteria that elicits an immune response in mammalian cells. The presence of structurally diverse lipid A types in the human gut bacteria has been suggested before, and this appears associated with the immune response. However, lipid A structures and their quantitative heterogeneity have not been well characterized. In this study, a method of analysis for lipid A using liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-QTOF/MS) was developed and applied to the analyses of Escherichia coli and Bacteroidetes strains. In general, phosphate compounds adsorb on stainless-steel piping and cause peak tailing, but the use of an ammonia-containing alkaline solvent produced sharp lipid A peaks with high sensitivity. The method was applied to E. coli strains, and revealed the accumulation of lipid A with abnormal acyl side chains in knockout strains as well as known diphosphoryl hexa-acylated lipid A in a wild-type strain. The analysis of nine representative strains of Bacteroidetes showed the presence of monophosphoryl penta-acylated lipid A characterized by a highly heterogeneous main acyl chain length. Comparison of the structures and amounts of lipid A among the strains suggested a relationship between lipid A profiles and the phylogenetic classification of the strains.

Sign in / Sign up

Export Citation Format

Share Document