Development of an off-line capillary column IMAC phosphopeptide enrichment method for label-free phosphorylation relative quantification

Choi, Hyunwoo; Lee, Sungjoon; Jun, Chang‐Duk; Park, Chul–Seung

doi:10.1016/j.jchromb.2011.08.035

Cited by 7 publications

(6 citation statements)

References 34 publications

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“…Resulting tryptic digest peptide samples were de-salted for subsequent phosphopeptide enrichment procedure. Phosphopeptides enriched with IMAC POROS 20 MC were subsequently subjected to the micro RPLC-MS/MS analysis described previously ( Choi et al, 2011 ). Briefly, analyses of phosphopeptide-enriched samples were performed using Agilent 1100 Series high-performance liquid chromatography (HPLC) (Agilent Technologies) coupled to a linear quadrupole ion trap mass spectrometer (LTQ, Thermo Finnigan, USA).…”

Section: Methodsmentioning

confidence: 99%

A Pair of Oviduct-Born Pickpocket Neurons Important for Egg-Laying in Drosophila melanogaster

Lee

Choi

Zhang

et al. 2016

Molecules and Cells

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During copulation, male Drosophila transfers Sex Peptide (SP) to females where it acts on internal sensory neurons expressing pickpocket (ppk). These neurons induce a post-mating response (PMR) that includes elevated egg-laying and refractoriness to re-mating. Exactly how ppk neurons regulate the different aspects of the PMR, however, remains unclear. Here, we identify a small subset of the ppk neurons which requires expression of a pre-mRNA splicing factor CG3542 for egg-laying, but not refractoriness to mating. We identify two CG3542-ppk expressing neurons that innervate the upper oviduct and appear to be responsible for normal egg-laying. Our results suggest specific subsets of the ppk neurons are responsible for each PMR component.

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Section: Methodsmentioning

confidence: 99%

A Pair of Oviduct-Born Pickpocket Neurons Important for Egg-Laying in Drosophila melanogaster

Lee

Choi

Zhang

et al. 2016

Molecules and Cells

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“…described elsewhere (Choi et al, 2011). Briefly, peptides were separated by C18 reversed phase capillary column and Agilent HP1100 quaternary LC pump.…”

Section: Micro-lc-ms/ms Analysismentioning

confidence: 99%

SNX14 is a bifunctional negative regulator for neuronal 5-HT6 receptor signaling

Park

Kim

et al. 2015

Journal of Cell Science

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The 5-hydroxytryptamine (5-HT, also known as serotonin) subtype 6 receptor (5-HT 6 R, also known as HTR6) plays roles in cognition, anxiety and learning and memory disorders, yet new details concerning its regulation remain poorly understood. In this study, we found that 5-HT 6 R directly interacted with SNX14 and that this interaction dramatically increased internalization and degradation of 5-HT 6 R. Knockdown of endogenous SNX14 had the opposite effect. SNX14 is highly expressed in the brain and contains a putative regulator of G-protein signaling (RGS) domain. Although its RGS domain was found to be non-functional as a GTPase activator for Gas, we found that it specifically bound to and sequestered Gas, thus inhibiting downstream cAMP production. We further found that protein kinase A (PKA)-mediated phosphorylation of SNX14 inhibited its binding to Gas and diverted SNX14 from Gas binding to 5-HT 6 R binding, thus facilitating the endocytic degradation of the receptor. Therefore, our results suggest that SNX14 is a dual endogenous negative regulator in 5-HT 6 R-mediated signaling pathway, modulating both signaling and trafficking of 5-HT 6 R.

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“…Enrichment of the phosphorylated peptides prior to mass spectrometric analysis is often required [3] . Calculating the phosphorus content of each phosphorylated peptide because of different retention times is difficult.…”

Section: Experimental Design Materials and Methodsmentioning

confidence: 99%

Data on the peptide mapping and MS identification for phosphorylated peptide

Wang

Liu

et al. 2016

Data in Brief

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This article contains peptides mapping, mass spectrometry and processed data related to the research “Identification and quantification of the phosphorylated ovalbumin by high resolution mass spectrometry under dry-heating treatment” [1]. Fourier transform ion cyclotron mass spectrometry (FTICR MS) was used to investigate the specific phosphorylation sites and the degree of phosphorylation (DSP) at each site. Specifically, phosphorylated peptides were monitored through mass shift on the FTICR MS spectrum. DSP was evaluated through the relative abundance levels of the FTICR MS spectrometry. From these data, the calculation method of DSP was exemplified.

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Development of an off-line capillary column IMAC phosphopeptide enrichment method for label-free phosphorylation relative quantification

Cited by 7 publications

References 34 publications

A Pair of Oviduct-Born Pickpocket Neurons Important for Egg-Laying in Drosophila melanogaster

A Pair of Oviduct-Born Pickpocket Neurons Important for Egg-Laying in Drosophila melanogaster

SNX14 is a bifunctional negative regulator for neuronal 5-HT6 receptor signaling

Data on the peptide mapping and MS identification for phosphorylated peptide

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