2012
DOI: 10.1590/s1415-47572012000200009
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Separomics applied to the proteomics and peptidomics of low-abundance proteins: choice of methods and challenges - a review

Abstract: The enrichment and isolation of proteins are considered limiting steps in proteomic studies. Identification of proteins whose expression is transient, those that are of low-abundance, and of natural peptides not described in databases, is still a great challenge. Plant extracts are in general complex, and contaminants interfere with the identification of proteins involved in important physiological processes, such as plant defense against pathogens. This review discusses the challenges and strategies of separo… Show more

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Cited by 13 publications
(12 citation statements)
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“…Among plant tissues, this problem is very apparent in leaves and other photosynthetic tissues, where the majority of proteins is highly hindered by a single protein, ribulose bisphosphate carboxylase (RuBisCo) (88).Therefore, a group of methodological strategies for the separation of protein molecules for proteomics, called separomics, is applied for the identification of low-abundance proteins and peptides in plants (89). Different techniques have been developed to remove or reduce a substantial portion of RuBisCo from leaf protein extracts, such as the partial removal of …”
Section: Cvjetko Et Al Proteomics Of Heavy Metal Toxicity In Plantsmentioning
confidence: 99%
“…Among plant tissues, this problem is very apparent in leaves and other photosynthetic tissues, where the majority of proteins is highly hindered by a single protein, ribulose bisphosphate carboxylase (RuBisCo) (88).Therefore, a group of methodological strategies for the separation of protein molecules for proteomics, called separomics, is applied for the identification of low-abundance proteins and peptides in plants (89). Different techniques have been developed to remove or reduce a substantial portion of RuBisCo from leaf protein extracts, such as the partial removal of …”
Section: Cvjetko Et Al Proteomics Of Heavy Metal Toxicity In Plantsmentioning
confidence: 99%
“…Identification of low-abundance proteins in green tissue that contains high amounts of polyphenols and many highly abundant proteins is one of the main challenges in banana membrane proteomics. Reduction in the level of high-abundance proteins in extracts prior to MS analysis has proved an efficient way to improve detection of low-abundance proteins ( Baracat et al, 2012 ). In addition, as a powerful tool for shotgun proteomics, isobaric tags for relative and absolute quantification (iTRAQ) have been widely applied in plant proteome analysis ( Kosová et al, 2011 ).…”
Section: Introductionmentioning
confidence: 99%
“… 213 , 230 Two main tools are used to isolate proteins: (1) the 2-dimensional electrophoresis (2-DE) associated with mass spectrometry (MS) and (2) liquid chromatography associated with MS, each one with its own limitations. 230 - 232 Obtaining native proteins is a challenge in proteomics or peptidomics, due to high protein complexity in samples, as the occurrence of post-translational modifications. Alternative strategies applied to extraction, purification, biochemical, and functional analyses of these molecules have been proposed, favoring access to structural and functional information of hard-to-reach proteins and peptides.…”
Section: Bioinformatic Tools and Databases For Plant Ampsmentioning
confidence: 99%