Two‐dimensional electrophoresis of human serum proteins modified by ampicillin during therapeutic treatment

Magi, Barbara; Marzocchi, Barbara; Bini, Luca; Cellesi, C; Rossolini, A; Pallini, Vitaliano

doi:10.1002/elps.11501601198

Cited by 15 publications

(10 citation statements)

References 17 publications

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“…Tissot et al demonstrated characteristic changes in the plasma 2-DE pattern indicative of monoclonal gammopathies, hypergammaglobulinemia, hepatic failure, chronic renal failure, and hemolytic anemia (42) as well as progressive changes during fetal development (43). In vivo covalent binding of ampicillin to multiple plasma proteins (not simply albumin) was demonstrated (44), providing a general method to probe covalent drug binding in plasma following drug treatment. Other changes have been reported associated with malnutrition (45), haptoglobin in Duchenne muscular dystrophy (46), haptoglobin in Down syndrome (47), apoA-I during parturition (48), return from extended space flight (49) (possibly an acute phase response to resuming 1 ϫ g), apoA-I isoforms in heart disease (50), human chorionic gonadotropin isoforms in patients with trophoblastic tumors (51), apoE isoforms in chronic spinal pain (52), and oxidized plasma proteins in Alzheimer's disease (53).…”

Section: Two-dimensional Electrophoresismentioning

confidence: 99%

The Human Plasma Proteome

Anderson¹,

Anderson²

2002

Molecular & Cellular Proteomics

3,864

1,585

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The human plasma proteome holds the promise of a revolution in disease diagnosis and therapeutic monitoring provided that major challenges in proteomics and related disciplines can be addressed. Plasma is not only the primary clinical specimen but also represents the largest and deepest version of the human proteome present in any sample: in addition to the classical "plasma proteins," it contains all tissue proteins (as leakage markers) plus very numerous distinct immunoglobulin sequences, and it has an extraordinary dynamic range in that more than 10 orders of magnitude in concentration separate albumin and the rarest proteins now measured clinically. Although the restricted dynamic range of conventional proteomic technology (two-dimensional gels and mass spectrometry) has limited its contribution to the list of 289 proteins (tabulated here) that have been reported in plasma to date, very recent advances in multidimensional survey techniques promise at least double this number in the near future. Abundant scientific evidence, from proteomics and other disciplines, suggests that among these are proteins whose abundances and structures change in ways indicative of many, if not most, human diseases. Nevertheless, only a handful of proteins are currently used in routine clinical diagnosis, and the rate of introduction of new protein tests approved by the United States Food and Drug Administration (FDA) has paradoxically declined over the last decade to less than one new protein diagnostic marker per year. We speculate on the reasons behind this large discrepancy between the expectations arising from proteomics and the realities of clinical diagnostics and suggest approaches by which protein-disease associations may be more effectively translated into diagnostic tools in the future.

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Section: Two-dimensional Electrophoresismentioning

confidence: 99%

The Human Plasma Proteome

Anderson¹,

Anderson²

2002

Molecular & Cellular Proteomics

3,864

1,585

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“…More recently, both MS and immunological approaches have been used by several groups. Immunological procedures allowed the identification of some serum proteins, including HSA and transferrin, as targets for haptenation by ampicillin [16]. Similarly, an anti-flucloxacillin antibody has been used to detect adducts formed in the liver of flucloxacillin-treated rats [17].…”

Section: Introductionmentioning

confidence: 99%

Study of Protein Haptenation by Amoxicillin Through the Use of a Biotinylated Antibiotic

et al. 2014

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Allergic reactions towards β-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a β-lactams, to bind covalently to proteins, in a process known as haptenation, is considered necessary for induction of a specific immunological response. Identification of the proteins modified by β-lactams and elucidation of the relevance of this process in allergic reactions requires sensitive tools. Here we describe the preparation and characterization of a biotinylated amoxicillin analog (AX-B) as a tool for the study of protein haptenation by amoxicillin (AX). AX-B, obtained by the inclusion of a biotin moiety at the lateral chain of AX, showed a chemical reactivity identical to AX. Covalent modification of proteins by AX-B was reduced by excess AX and vice versa, suggesting competition for binding to the same targets. From an immunological point of view, AX and AX-B behaved similarly in RAST inhibition studies with sera of patients with non-selective allergy towards β-lactams, whereas, as expected, competition by AX-B was poorer with sera of AX-selective patients, which recognize AX lateral chain. Use of AX-B followed by biotin detection allowed the observation of human serum albumin (HSA) modification by concentrations 100-fold lower that when using AX followed by immunological detection. Incubation of human serum with AX-B led to the haptenation of all of the previously identified major AX targets. In addition, some new targets could be detected. Interestingly, AX-B allowed the detection of intracellular protein adducts, which showed a cell type-specific pattern. This opens the possibility of following the formation and fate of AX-B adducts in cells. Thus, AX-B may constitute a valuable tool for the identification of AX targets with high sensitivity as well as for the elucidation of the mechanisms involved in allergy towards β-lactams.

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“…MS is a sensitive and accurate method for the determination of PTMs and, thus, for the study of protein haptenation. Using two-dimensional electrophoresis and immunological detection, HSA and transferrin were identified as penicilloyl-modified proteins in plasma from ampicillin-treated patients [42]. More recently, the modification of HSA by flucloxacillin has been studied in vitro and in tolerant patients by LC-MS/MS, and Lys 190 and 212 of HSA have been identified as the main sites for addition of this antibiotic [43].…”

Section: Applications Of Proteomics To the Study Of Immunological Reamentioning

confidence: 98%

Proteomics in immunological reactions to drugs

Ariza

Montañez

Pérez‐Sala

2011

Current Opinion in Allergy &Amp; Clinical Immunology

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Immunological reactions to drugs pose important clinical problems. Since early works exploring drug-protein interactions, there has been steady progress in this field. However, the mechanisms involved remain incompletely understood. The availability of proteomic techniques with high resolution and sensitivity presents a unique opportunity to tackle this subject from a broad perspective, integrating work in model systems and in patients. Chemical and metabolic characterization of immunological reactions to drugs may also help in the prevention, diagnosis and/or treatment of these processes.

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Two‐dimensional electrophoresis of human serum proteins modified by ampicillin during therapeutic treatment

Cited by 15 publications

References 17 publications

The Human Plasma Proteome

The Human Plasma Proteome

Study of Protein Haptenation by Amoxicillin Through the Use of a Biotinylated Antibiotic

Proteomics in immunological reactions to drugs

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