Carotid atherosclerotic plaques: Proteomics study after a low‐abundance protein enrichment step

Abstract

Atherosclerosis is one of the most important causes of cardiovascular and cerebrovascular events. Although phenotypic differentiation between stable and unstable plaques is currently possible, proteomic analysis of the atherosclerotic plaque could offer a global view of the atherosclerosis pathology. With the objective to highlight the detection of low-abundance proteins, we reduced the dynamic range of proteins by combinatorial peptide ligand library treatment of human carotid artery atherosclerotic plaques. After enrichment step, abundance of major proteins was decreased, revealing different protein profiles as assessed by both SDS-polyacrylamide gel electrophoresis and two-dimensional electrophoresis comparative analyses. Identification of proteins that were contained in a spot allowed finding large differences between noncomplicated and complicated plaques from carotid atherosclerotic lesions. Novel low-abundance proteins were detected correlating very well with biological alterations related to atherosclerosis (heat shock protein 27 (HSP27) isoforms, aldehyde dehydrogenase, moesin, Protein kinase C delta-binding protein, and inter-α trypsin inhibitor family heavy chain-related protein (ITIH4)). At the same time, the differential expression of known proteins of interest such as hemoglobin β-chain and heat shock protein 27 between noncomplicated and hemorrhagic complicated plaques was maintained after enrichment step. The detection of different isoforms of a low-abundance protein such as heat shock protein 27 species was actually improved after enrichment of tissue protein extracts. All of these findings clearly support further investigations in view to confirm the role of these proteins as possible biomarkers.

Proteomic analysis and comparison of intra- and extracranial cerebral atherosclerosis responses to hyperlipidemia in rabbits

“…Therefore, HSP70 may have AS-protective properties via anti-inflammatory underlying mechanisms in ICAS compared with ECAS. This finding is consistent with a previous study, which examined the proteomic analysis of stable and unstable carotid atherosclerotic plaques (6). The findings of the present study may partly explain previous contradictory observations on the differences in cerebral AS mechanisms.…”

“…Rabbits were randomly divided into two groups: Control (A; n=9) and high-fat (B; n=9). Rabbits in group A continued to receive regular rabbit feed and group B were fed a high-fat diet (2% cholesterol, 6% peanut oil and 92% basic feed) for 12 weeks based on the methods of previous studies (6,9,10). Rabbits in groups A and B were fed restricted diets with an equal amount of food (150–200 g per day) and free access to water.…”

“…The role of hyperlipidemia in ICAS and ECAS remains unclear due to controversial previously reported findings. A previous study conducted proteomics analyses of the common carotid artery by obtaining human carotid atherosclerotic plaques, and demonstrated certain proteins are in low-abundance, including heat shock protein 27 (HSP27) isoforms, aldehyde dehydrogenase, moesin, protein kinase C δ-binding protein and inter-α-trypsin inhibitor family heavy chain-related protein are correlated with biological alterations associated with atherosclerosis (6). In our previous study, HSP70 expression levels were demonstrated to vary between ICAS and ECAS; however, the effect of other protein expressions on ICAS and ECAS remains unclear (7).…”

Liquid phase based separation systems for depletion, prefractionation, and enrichment of proteins in biological fluids and matrices for in‐depth proteomics analysis—An update covering the period 2011–2014

“…Malaud et al [34] reduced the dynamic range of proteins by CPLL treatment of humancarotid artery atherosclerotic plaques. After the enrichment step, the abundance of major proteins was decreased, revealing different protein profiles as assessed by both SDS-PAGE and 2-DE comparative analyses and then identification of protein using LC-MS/MS.…”

Mark Twain: How to fathom the depth of your pet proteome

“…Currently, by treating with the ProteoMiner beads 150 mL of urines, we can detect, in a single sweep, about 1800 unique gene products (work in progress). While used for the detection of additional proteins and isoforms that are normally not detectable prior to CPLL treatment, enlarging thus the knowledge of proteomes, the 2D results have been used for the detection of expression differences along with identification [25]. In this last case a grand total of 3300 spots could be counted.…”