Abstract

Protein ladder sequencing is a new technique developed in our laboratory to sequence polypeptides including those with modified amino acids. One of the important applications of this technique is the study of protein phosphorylation. To understand the specificities of different protein kinases and to further explore the potentials of ladder sequencing, a model peptide, which contains multiple serine-phosphorylation sites, was synthesized chemically. This peptide is being used to study the specificities of several different protein kinases and the stochiometries of phosphorylation. In addition, variously phosphorylated (site and number) model peptides (same sequence as above peptide) will be synthesized to examine whether our mass spectrometric technique can be used to obtain detailed information concerning the stochiometry of phosphorylation. A paper describing these results is in preparation.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-23
Application #
5221598
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
23
Fiscal Year
1996
Total Cost
Indirect Cost

  Publications

Manning, Lois R; Popowicz, Anthony M; Padovan, Julio C et al. (2017) Gel filtration of dilute human embryonic hemoglobins reveals basis for their increased oxygen binding. Anal Biochem 519:38-41
Boice, Michael; Salloum, Darin; Mourcin, Frederic et al. (2016) Loss of the HVEM Tumor Suppressor in Lymphoma and Restoration by Modified CAR-T Cells. Cell 167:405-418.e13
Chait, Brian T; Cadene, Martine; Olinares, Paul Dominic et al. (2016) Revealing Higher Order Protein Structure Using Mass Spectrometry. J Am Soc Mass Spectrom 27:952-65
Krutchinsky, Andrew N; Padovan, Júlio C; Cohen, Herbert et al. (2015) Maximizing ion transmission from atmospheric pressure into the vacuum of mass spectrometers with a novel electrospray interface. J Am Soc Mass Spectrom 26:649-58
Mast, Fred D; Rachubinski, Richard A; Aitchison, John D (2015) Signaling dynamics and peroxisomes. Curr Opin Cell Biol 35:131-6
Krutchinsky, Andrew N; Padovan, Júlio C; Cohen, Herbert et al. (2015) Optimizing electrospray interfaces using slowly diverging conical duct (ConDuct) electrodes. J Am Soc Mass Spectrom 26:659-67
Oricchio, Elisa; Papapetrou, Eirini P; Lafaille, Fabien et al. (2014) A cell engineering strategy to enhance the safety of stem cell therapies. Cell Rep 8:1677-1685
Zhong, Yu; Morris, Deanna H; Jin, Lin et al. (2014) Nrbf2 protein suppresses autophagy by modulating Atg14L protein-containing Beclin 1-Vps34 complex architecture and reducing intracellular phosphatidylinositol-3 phosphate levels. J Biol Chem 289:26021-37
Mathur, Aabhas; Blais, Steven; Goparaju, Chandra M V et al. (2013) Development of a biosensor for detection of pleural mesothelioma cancer biomarker using surface imprinting. PLoS One 8:e57681
Peterson, Shaun E; Li, Yinyin; Wu-Baer, Foon et al. (2013) Activation of DSB processing requires phosphorylation of CtIP by ATR. Mol Cell 49:657-67

Showing the most recent 10 out of 67 publications