Two‐dimensional electrophoresis of human placental mitochondria and protein identification by mass spectrometry: Toward a human mitochondrial proteome

Rabilloud, Thierry; Kieffer, Sylvie; Procaccio, Vincent; Louwagie, Mathilde; Courchesne, Paul; Patterson, Scott D.; Martinez, Pascal; Garin, Jérôme; Lunardi, Joël

doi:10.1002/elps.1150190616

Cited by 206 publications

(169 citation statements)

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“…This latter approach adds valuable localization and functional information and is therefore quite attractive. One of the first example of such subcellular proteomes has been made on mitochondria [6], but other, more recent examples have been shown on lysosomes [7], golgi [8] or chloroplasts [9].…”

Section: Introductionmentioning

confidence: 99%

Progress in the definition of a reference human mitochondrial proteome

et al. 2003

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Owing to the complexity of higher eukaryotic cells, a complete proteome is likely to be very difficult to achieve. However, advantage can be taken of the cell compartmentalization to build organelle proteomes, which can moreover be viewed as specialized tools to study specifically the biology and "physiology" of the target organelle. Within this frame, we report here the construction of the human mitochondrial proteome, using placenta as the source tissue. Protein identification was carried out mainly by peptide mass fingerprinting. The optimization steps in two-dimensional electrophoresis needed for proteome research are discussed. However, the relative paucity of data concerning mitochondrial proteins is still the major limiting factor in building the corresponding proteome, which should be a useful tool for researchers working on human mitochondria and their deficiencies.Comment: website publisher http://www.interscience.wiley.co

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Section: Introductionmentioning

confidence: 99%

Progress in the definition of a reference human mitochondrial proteome

et al. 2003

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“…The mitochondrial inner membrane is of particular interest because it contains proteins of the respiratory chain as well as many carrier proteins and ion channels that certainly play a key role in mitochondrial function and cell homeostasis. Over the past few years, a number of approaches, such as two-dimensional PAGE and more recently one-dimensional PAGE, have allowed partial characterization of the proteome of entire mitochondria from various species (3)(4)(5)(6). However, our knowledge of the protein composition of the mitochondrial inner membrane is still largely incomplete.…”

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Proteomic Analysis of the Mouse Liver Mitochondrial Inner Membrane

Cruz

Xenarios

Langridge³

et al. 2003

Journal of Biological Chemistry

229

157

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Mitochondria play a crucial role in cellular homeostasis, which justifies the increasing interest in mapping the different components of these organelles. Here we have focused our study on the identification of proteins of the mitochondrial inner membrane (MIM). This membrane is of particular interest because, besides the well known components of the respiratory chain complexes, it contains several ion channels and many carrier proteins that certainly play a key role in mitochondrial function and, therefore, deserve to be identified at the molecular level. To achieve this goal we have used a novel approach combining the use of highly purified mouse liver mitochondrial inner membranes, extraction of membrane proteins with organic acid, and two-dimensional liquid chromatography coupled to tandem mass spectrometry. This procedure allowed us to identify 182 proteins that are involved in several biochemical processes, such as the electron transport machinery, the protein import machinery, protein synthesis, lipid metabolism, and ion or substrate transport. The full range of isoelectric point (3.9 -12.5), molecular mass (6 -527 kDa), and hydrophobicity values (up to 16 transmembrane predicted domains) were represented. In addition, of the 182 proteins found, 20 were unknown or had never previously been associated with the MIM. Overexpression of some of these proteins in mammalian cells confirmed their mitochondrial localization and resulted in severe remodeling of the mitochondrial network. This study provides the first proteome of the MIM and provides a basis for a more detailed study of the newly characterized proteins of this membrane.

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“…While this strategy can give you additional information about the subcellular localization of given set of proteins, it further expands the necessary labor if one's goal is to assay the total complement of cellular proteins. Even after enrichment, the previously mentioned difficulty of analyzing membrane proteins is present (11), and that subset of membrane proteins, which can be visualized, are often so diffuse on the gel that their subsequent identification is impossible.…”

Section: Proteomic Techniques: Avoiding the Gelmentioning

confidence: 99%

Proteomic Tools for Cell Biology

McDonald

Yates

2000

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Acquisition of large bodies of genomic sequence is facilitating the use of global techniques to assay cellular function. DNA microarrays have enabled the measurement of global mRNA levels and are able to detect changes in gene expression between different cellular states. Since much of the regulation of physiolgical processes happens post-translationally, measuring only the mRNA levels gives an incomplete picture. Strategies to assay global expression, localization, or interaction of proteins fall into the emerging field of proteomics, with various combinations of techniques being utilized to separate and identify proteins. In this review, we will present a general overview of the currently available proteomic tools and then give examples of how these tools are being utilized to answer questions in cell biology.

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Two‐dimensional electrophoresis of human placental mitochondria and protein identification by mass spectrometry: Toward a human mitochondrial proteome

Cited by 206 publications

References 46 publications

Progress in the definition of a reference human mitochondrial proteome

Progress in the definition of a reference human mitochondrial proteome

Proteomic Analysis of the Mouse Liver Mitochondrial Inner Membrane

Proteomic Tools for Cell Biology

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