Proteomic evidence for the plasticity of cultured vascular smooth muscle cells

Abstract

Morphological, functional, and gene expression studies have established the phenotypic plasticity of smooth muscle cells (SMCs). These cells have been shown to respond to environmental stimulants such as extracellular matrix and growth factors. Cellular changes can vary between extremes defined as the contractile and synthetic states. Various growth factors have been shown to have profound effects on the phenotype of these cells. In this study, we intended to investigate the effects of growth factor-rich medium on the protein expression of vascular SMCs in culture. Interestingly, transiently changing the type of medium did not result in any apparent morphological differences, yet we hypothesized that some cellular factors might still be altered. In order to understand what kind of intracellular molecular changes should be expected to occur during the medium change, we analyzed global protein expression changes using nano-LC-MS/MS in smooth muscle cell cultures that were isolated and grown in one medium formulation and temporarily switched to the other. Our data indicate that proteins playing a role in energy metabolism (glycolysis), translation, and folding of proteins are affected, along with regulatory molecules and cytoskeletal proteins. The individual proteins and their significance are discussed within the scope of this paper.

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“…× 250 mm, Waters) at a flow rate of 300 nl/min, with a gradient of 5–40% mobile phase B (0.1 formic acid in hyper grade acetonitrile, Merck), over a 90-min period. The MS parameters were set as reported previously [9]. The instrument was run in positive ion V mode, applying the MS and MS/MS functions over 1.5-s intervals with 6-V low-energy and 15- to 40-V high energy collisions.…”

Biochemical and Proteomic Analysis of a Potential Anticancer Agent: Palladium(II) Saccharinate Complex of Terpyridine Acting through Double Strand Break Formation

“…Four independent plates of untreated cells and three independent biological repeats of treated cells were prepared for this analysis. A modified protocol from ref was used for sample preparation. Briefly, cells were lysed with UPX extraction buffer (Expedeon) following two washes with ice cold 50 mM ammonium bicarbonate solution.…”

Photocatalytically Active Graphitic Carbon Nitride as an Effective and Safe 2D Material for In Vitro and In Vivo Photodynamic Therapy

“…The samples were processed using the Acquity sample manager with mobile phase consisting of A1 (0.1% formic acid in LC/MS grade water) and B1 (0.1% formic acid in hyper grade acetonitrile) solutions with a sample flow rate of 300 nL min −1 over 90 min. The MS settings were used as previously described . Data was acquired in a range of m / z 50–1600 Da.…”

“…The MS settings were used as previously described. [49] Data was acquired in a range of m/z 50-1600 Da. The instrument was run in positive ion V mode with a scan time of 1.5 s, with 6 V low energy and 15-40 V high energy collisions.…”