Profiling of alterations in platelet proteins during storage of platelet concentrates

Thiele, Thomas; Steil, Leif; Gebhard, Simon; Scheffold, Christian; Hammer, Elke; Brigulla, Matthias; Lübenow, Norbert; Clemetson, Kenneth J.; Völker, Uwe; Greinacher, Andreas

doi:10.1111/j.1537-2995.2007.01255.x

Cited by 97 publications

(98 citation statements)

References 32 publications

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“…3 The few storage studies suggested apoptosis and cytoskeleton as affected systems that may be associated with shortened survival of transfused platelets. [4][5][6] Whereas some changes were attributed to active translation from preexisting messenger RNAs, 7 others were due to altered protein posttranslational modifications (PTMs), such as phosphorylation 8,9 and proteolysis. 4,8 However, despite the importance of PTMs to protein and platelet function, the occurrence and regulation of most PTMs during platelet storage remains unknown.…”

Section: Introductionmentioning

confidence: 99%

“…[4][5][6] Whereas some changes were attributed to active translation from preexisting messenger RNAs, 7 others were due to altered protein posttranslational modifications (PTMs), such as phosphorylation 8,9 and proteolysis. 4,8 However, despite the importance of PTMs to protein and platelet function, the occurrence and regulation of most PTMs during platelet storage remains unknown. Proteolysis represents one of the important, but often overlooked, PTMs that is ubiquitous, irreversible, and affects every protein in the cell through excision of the initiator methionine, removal of signal peptides or propeptides, and eventually degradation.…”

Section: Introductionmentioning

confidence: 99%

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TAILS N-terminomics of human platelets reveals pervasive metalloproteinase-dependent proteolytic processing in storage

Prudova

Serrano

Eckhard

et al. 2014

Blood

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Key Points• TAILS proteomics identified 2938 human platelet proteins, pervasive proteolytic processing, and precise proteolytic cleavage sites in stored platelets.• During storage, metalloproteinases were predominantly involved in proteolytic processing, while other proteinases were mainly involved in degradation.Proteases, and specifically metalloproteinases, have been linked to the loss of platelet function during storage before transfusion, but the underlying mechanisms remain unknown. We used a dedicated N-terminomics technique, iTRAQ terminal amine isotopic labeling of substrates (TAILS), to characterize the human platelet N-terminome, proteome, and posttranslational modifications throughout platelet storage over 9 days under blood-banking conditions. From the identified 2938 proteins and 7503 unique peptides, we characterized N-terminal methionine excision, co-and posttranslational Na acetylation, protein maturation, and proteolytic processing of proteins in human platelets. We also identified for the first time 10 proteins previously classified by the Human Proteome Organization as "missing" in the human proteome. Most N termini (77%) were internal neo-N termini (105 were novel potential alternative translation start sites, and 2180 represented stable proteolytic products), thus highlighting a prominent yet previously uncharacterized role of proteolytic processing during platelet storage. Protease inhibitor studies revealed metalloproteinases as being primarily responsible for proteolytic processing (as opposed to degradation) during storage. System-wide identification of metalloproteinase and other proteinase substrates and their respective cleavage sites suggests novel mechanisms of the effect of proteases on protein activity and platelet function during storage. All data sets and metadata are available through ProteomeXchange with the data set identifier PXD000906. (Blood. 2014;124(26):e49-e60)

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

TAILS N-terminomics of human platelets reveals pervasive metalloproteinase-dependent proteolytic processing in storage

Prudova

Serrano

Eckhard

et al. 2014

Blood

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“…Protein Identification by Mass Spectrometry-Silver nitrate-stained gel slices were destained by incubation in 30 mM K 3 [Fe(CN) 6 ], 100 mM Na 2 S 2 O 3 until colorless and washed three times in water before processing gel slices as described previously (29). Briefly gel pieces were washed twice with 200 l of 20 mM NH 4 HCO 3 , 50% (v/v) ACN for 30 min at 37°C and dried by adding 200 l of ACN two times for 15 min.…”

Section: Methodsmentioning

confidence: 99%

Novel Activities of Glycolytic Enzymes in Bacillus subtilis

Commichau

Rothe

Herzberg

et al. 2009

Molecular & Cellular Proteomics

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Glycolysis is one of the most important metabolic pathways in heterotrophic organisms. Several genes encoding glycolytic enzymes are essential in many bacteria even under conditions when neither glycolytic nor gluconeogenic activities are required. In this study, a screening for in vivo interaction partners of glycolytic enzymes of the soil bacterium Bacillus subtilis was used to provide a rationale for essentiality of glycolytic enzymes. Glycolytic enzymes proved to be in close contact with several other proteins, among them a high proportion of essential proteins. Among these essential interaction partners, other glycolytic enzymes were most prominent. Two-hybrid studies confirmed interactions of phosphofructokinase with phosphoglyceromutase and enolase. Such a complex of glycolytic enzymes might allow direct substrate channeling of glycolytic intermediates. Moreover we found associations of glycolytic enzymes with several proteins known or suspected to be involved in RNA processing and degradation. One of these proteins, Rny (YmdA), which has so far not been functionally characterized, is required for the processing of the mRNA of the glycolytic gapA operon. Two-hybrid analyses confirmed the interactions between the glycolytic enzymes phosphofructokinase and enolase and the enzymes involved in RNA processing, RNase J1, Rny, and polynucleotide phosphorylase. Moreover RNase J1 interacts with its homologue RNase J2. We suggest that this complex of mRNA processing and glycolytic enzymes is the B. subtilis equivalent of the RNA degradosome. Our findings suggest that the functional interaction of glycolytic enzymes with essential proteins may be the reason why they are indispensable. Glycolysis is a central metabolic pathway that appeared early in the evolution of life (1). Major functions of the glycolytic pathway are the generation of precursors for anabolic reactions and the conservation of energy that is needed to fuel all other cellular processes. The glycolytic pathway is conventionally divided into two parts: (i) the upper part, also referred to as the preparatory phase of glycolysis because the reactions of this part consume energy to convert the incoming sugars to triose phosphates, and (ii), the lower part or payoff phase that is characterized by the net gain of energy and the formation of reduction equivalents. Although glycolysis is highly conserved from archaea and bacteria to man not all organisms use this pathway for the oxidization of glucose. Escherichia coli is able to oxidize glucose via glycolysis, but the Entner-Doudoroff and the pentose phosphate pathways may replace the preparatory phase. In contrast to E. coli, the Entner-Doudoroff pathway is not present in Bacillus subtilis (2). Interestingly the enzymes of the upper glycolytic part seem to be completely absent in many archaea (3, 4). These few examples show that there is a high plasticity in how archaea and bacteria can feed glucose into the triose phosphate part of glycolysis. This plasticity is in good agreement with the observation that...

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“…Thus, 2D-DIGE has emerged as a powerful tool to comprehensively assess changes in the cytoplasmic platelet proteome while minimizing interassay variability. 14,15 Using 2D-DIGE and mass spectrometry, we here determined that specific activation of platelet GPVI results in a significant change in abundance of several proteins. Consecutively, we also analyzed functional changes of 2 identified proteins and their relevance for platelet aggregation and blood coagulation.…”

Section: Introductionmentioning

confidence: 99%

Identification of novel downstream targets of platelet glycoprotein VI activation by differential proteome analysis: implications for thrombus formation

Schulz

Leuschen

Fröhlich³

et al. 2010

Blood

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Platelets play a key role in hemostasis and various diseases including arterial thrombosis. Glycoprotein VI (GPVI) mediates adhesion to collagen structures exposed at sites of vascular injury and subsequent platelet activation. We determined the effects of specific activation of GPVI on the human platelet proteome. Isolated human platelets were stimulated with an activating monoclonal antibody specific for GPVI. Platelet proteins were analyzed by 2-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry. We identified 8 differentially abundant proteins associated with cell signaling, metabolism, organization and rearrangement of the cytoskeleton, and membrane trafficking. Differentially abundant proteins included aldose reductase (AR), beta-centractin, charged multivesicular body protein 3, Src substrate cortactin, ERp57, and pleckstrin. Importantly, GPVI-modulated protein abundance was functionally relevant. Correspondingly, AR enzyme activity significantly increased upon GPVI activation and inhibition of AR resulted in reduced platelet aggregation. Furthermore, ERp57 was released upon ligation of platelet GPVI and increased the activity of tissue factor, a major initiator of blood coagulation. In summary, GPVI activation results in differential changes in abundance of platelet proteins, including AR and ERp57, which support platelet aggregation and plateletdependent coagulation. These results provide further insight into the mechanisms that underlie platelet activation through the GPVI receptor and may help to identify novel pharmacologic targets.

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Profiling of alterations in platelet proteins during storage of platelet concentrates

Abstract: DIGE is a tool for comprehensively assessing the impact of storage on the global proteome profile of therapeutic PCs. Most of the changes detected are in high-abundance PLT proteins.

Cited by 97 publications

References 32 publications

TAILS N-terminomics of human platelets reveals pervasive metalloproteinase-dependent proteolytic processing in storage

TAILS N-terminomics of human platelets reveals pervasive metalloproteinase-dependent proteolytic processing in storage

Novel Activities of Glycolytic Enzymes in Bacillus subtilis

Identification of novel downstream targets of platelet glycoprotein VI activation by differential proteome analysis: implications for thrombus formation

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