Abstract
For many years, HUVEC.com1 public database provides biological data relative to the proteome of human umbilical vein endothelial cells (HU-VECs), which are the most used human endothelial cell model in vascular biology. The proteins were identified using two-dimensional gel electrophoresis (2-DGE) for protein separation coupled with Matrix Assisted Laser Desorption-Ionization Mass Spectrometry (MALDI-TOF-MS) for identification. We present here an important update of HUVEC.com with 521 protein identifications as determined using Fourier transformed ion cyclotron resonance-mass spectrometry (FTICR-MS) applied to an unstained 2-DGE gel cut in 221 squared pieces; each identified protein being accompanied by a semi-quantitative three dimensional visualization is called “score imaging”. The squared analyzed gel and the alphabetical list of identified proteins, linked with their corresponding three-dimensional score imaging, are available at www.huvec.com. This original approach led to the establishment of the most protein-rich and informative database for HUVECs, as well as to the identification of some protein species, in particular with phosphorylation.
Highlights
From 2004, HUVEC.com shared a public database relative to human umbilical vein endothelial cells (HUVECs) proteome as assessed by the classical peptide mass fingerprinting approach combining two-dimensional gel electrophoresis (2-DGE) and Matrix Assisted Laser Desorption-Ionization Mass Spectrometry (MALDI-TOF-MS) [1]
In the goal to further enrich HUVEC.com, we used Fourier transformed ion cyclotron resonance-mass spectrometry (FTICR-MS) applied to an unstained 2. Two-Dimensional Gel Electrophoresis (2-DGE) gel cut in 221 equal rectangles to avoid the relatively poor sensitivity and the spot overlapping inherent to 2-DGE with classical staining [4,5]
Two identical gels were prepared: the first gel was stained successively with Colloidal Coomassie Blue (CCB) with silver nitrate; the second gel was cut in 221 equal rectangles without staining, the resulting grid pattern being matched against the stained control gel (Figure 1)
Summary
From 2004, HUVEC.com (www.huvec.com) shared a public database relative to human umbilical vein endothelial cells (HUVECs) proteome as assessed by the classical peptide mass fingerprinting approach combining two-dimensional gel electrophoresis (2-DGE) and Matrix Assisted Laser Desorption-Ionization Mass Spectrometry (MALDI-TOF-MS) [1]. Encountering a good audience with more than 100,000 visits till September 2012, HUVEC.com database appears as notably insufficient especially because of being restricted to a relatively low number of major, mainly soluble endothelial proteins. In the goal to further enrich HUVEC.com, we used Fourier transformed ion cyclotron resonance-mass spectrometry (FTICR-MS) applied to an unstained 2-DGE gel cut in 221 equal rectangles to avoid the relatively poor sensitivity and the spot overlapping inherent to 2-DGE with classical staining [4,5]. This study allowed for identifying some protein species in HUVECs, such as heat-shock proteins and proteins from the cytoskeleton
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