Amino acid sequence determination of phosphoenkephalins using liquid secondary ionization mass spectrometry

Dass, Chhabil; Mahalakshmi, P.

doi:10.1002/rcm.1290091213

Cited by 13 publications

(8 citation statements)

References 26 publications

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“…Several researchers have noted losses of HPO 3 from protonated phosphotyrosine peptides in low energy CID or PSD spectra [5,[7][8][9]12,14]. However these peptides all contained an arginine or lysine in their sequences.…”

Section: Resultsmentioning

confidence: 99%

Fragmentation of cationized phosphotyrosine containing peptides by atmospheric pressure MALDI/Ion trap mass spectrometry

Moyer

VonSeggern

Cotter

2003

J. Am. Soc. Mass Spectrom.

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An investigation of phosphate loss from sodium-cationized phosphotyrosine containing peptide ions was conducted using liquid infrared (2.94 m) atmospheric pressure matrixassisted laser desorption/ionization (AP MALDI) coupled to an ion trap mass spectrometer (ITMS). Previous experiments in our laboratory explored the fragmentation patterns of protonated phosphotyrosine containing peptides, which experience a loss of 98 Da under CID conditions in the ITMS. This loss of 98 Da is unexpected for phosphotyrosine, given the structure of its side chain. Phosphate loss from phosphotyrosine residues seems to be dependent on the presence of arginine or lysine residues in the peptide sequence. In the absence of a basic residue, the protonated phosphotyrosine peptides do not undergo losses of HPO 3 (⌬ 80 Da) nor HPO 3 ϩ H 2 O (⌬ 98 Da) in their CID spectra. However, sodium cationized phosphotyrosine containing peptides that do not contain arginine or lysine residues within their sequences do undergo losses of HPO 3 (⌬ 80 Da) and HPO 3 ϩ H 2 O (⌬ 98 Da) in their CID spectra.

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Section: Resultsmentioning

confidence: 99%

Fragmentation of cationized phosphotyrosine containing peptides by atmospheric pressure MALDI/Ion trap mass spectrometry

Moyer

VonSeggern

Cotter

2003

J. Am. Soc. Mass Spectrom.

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“…Activities of the neuropeptides may be altered by disulfide bond formation, glycosylation, COOH-terminal α-amidation, phosphorylaton, sulfation, and acetylation (12,(29)(30)(31)(32)(33)(34). This article, however, will focus on protease mechanisms for neuropeptide biosynthesis.…”

Section: Proteolytic Processing Of Proneuropeptidesmentioning

confidence: 99%

Proteases for Processing Proneuropeptides into Peptide Neurotransmitters and Hormones

Hook

Funkelstein

et al. 2008

Annu. Rev. Pharmacol. Toxicol.

220

304

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Summary Points List:1. Proteases are essential for proteolytic processing of proneuropeptide precursors into active peptide neurotransmitters and hormones.2. Secretory vesicles represent the primary subcellular site of neuropeptide biosynthesis, which are produced, stored, and secreted to mediate cell-cell communication. 3.Protease pathways for proneuropeptide processing have been elucidated consisting of (a) the newly identified cysteine protease cathepsin L with aminopeptidase B in secretory vesicles, and (b) the well-established, proprotein convertase family that include the neuroendocrine-specific prohormone convertases 1 and 2 (PC1/3 and PC2) with carboxypeptidase E. 4.Protease gene knockout experiments have validated the roles of PC1/3, PC2, as well as cathepsin L for the production of neuropeptides in nervous and endocrine tissues. 5.Endogenous regulators consisting of inhibitors and activators participate in the in vivo control of processing enzyme functions.6. Structural biology of protease and proneuropeptides will be important to understand interacting mechanisms for proneuropeptide processing. 7.Neuropeptidomics has recently been applied to investigations of neuropeptide systems for their primary sequence and structural identification, as well as quantitation by LC-MS/MS tandem mass spectrometry. 8.Proteomic studies have revealed functional protein families that participate in secretory vesicle functions for the production, storage, and secretion of neuropeptides.9. Pharmacological evaluation of unique specificities among neuropeptide processing systems will be valuable for design of future strategies to develop selective small molecule modulators of processing enzymes for therapeutic applications in health and disease.Future Issues: Areas of Neuropeptide Research for Exploration. 1.How are cathepsin L and prohormone convertase protease pathways coordinately regulated? 2.What is the proteolytic basis for tissue-specific processing of proneuropeptides, such as that for POMC?3. Selective and potent inhibitors of protease components for processing prohormones should be developed to facilitate basic and pharmacological research. 4.What are the structural features of prohormone and protease interactions for functional processing? Peptide neurotransmitters and peptide hormones, collectively known as neuropeptides, are required for cell-cell communication in neurotransmission and for regulation of endocrine functions. Neuropeptides are synthesized from protein precursors (termed proneuropeptides or prohormones) that require proteolytic processing primarily within secretory vesicles that store and secrete the mature neuropeptides to control target cellular and organ systems. This review describes interdisciplinary strategies that have elucidated two primary protease pathways for prohormone processing consisting of the cysteine protease pathway mediated by secretory vesicle cathepsin L and the well-known subtilisin-like proprotein convertase pathway that together support neuropeptide biosynthesis. Importantly...

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“…The detection of phosphopeptides can be achieved using either negative or positive ion modes. Several groups have successfully reported the use of negative ion ionization of phosphopeptides (Dass & Mahalakshmi, 1995;Annan et al, 2001) and their identification from complex cell extracts (Kocher, Allmaier, & Wilm, 2003;Old et al, 2009). However, the unpredictable nature of negative ion fragmentation and the limited sequence information obtainable from the dissociation of peptide anions make large-scale analyses very challenging.…”

Section: Phosphopeptide Identification By Mass Spectrometrymentioning

confidence: 99%

Subcellular phosphoproteomics

Trost

Bridon

Desjardins

et al. 2010

Mass Spectrometry Reviews

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Protein phosphorylation represents one of the most extensively studied post-translational modifications, primarily due to the emergence of sensitive methods enabling the detection of this modification both in vitro and in vivo. The availability of enrichment methods combined with sensitive mass spectrometry instrumentation has played a crucial role in uncovering the dynamic changes and the large expanding repertoire of this reversible modification. The structural changes imparted by the phosphorylation of specific residues afford exquisite mechanisms for the regulation of protein functions by modulating new binding sites on scaffold proteins or by abrogating protein-protein interactions. However, the dynamic interplay of protein phosphorylation is not occurring randomly within the cell but is rather finely orchestrated by specific kinases and phosphatases that are unevenly distributed across subcellular compartments. This spatial separation not only regulates protein phosphorylation but can also control the activity of other enzymes and the transfer of other post-translational modifications. While numerous large-scale phosphoproteomics studies highlighted the extent and diversity of phosphoproteins present in total cell lysates, the further understanding of their regulation and biological activities require a spatio-temporal resolution only achievable through subcellular fractionation. This review presents a first account of the emerging field of subcellular phosphoproteomics where cell fractionation approaches are combined with sensitive mass spectrometry methods to facilitate the identification of low abundance proteins and to unravel the intricate regulation of protein phosphorylation.

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Amino acid sequence determination of phosphoenkephalins using liquid secondary ionization mass spectrometry

Cited by 13 publications

References 26 publications

Fragmentation of cationized phosphotyrosine containing peptides by atmospheric pressure MALDI/Ion trap mass spectrometry

Fragmentation of cationized phosphotyrosine containing peptides by atmospheric pressure MALDI/Ion trap mass spectrometry

Proteases for Processing Proneuropeptides into Peptide Neurotransmitters and Hormones

Subcellular phosphoproteomics

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