Abstract

An enzymatic approach was developed to study the solution structure of the transcription factor MAX - a member of the basic-helix-loop-helix zipper family of DNA-binding proteins. This approach involves a proteolytic protection assay of the Max protein both in the absence and presence of DNA. The assay is based on the notion that residues of the protein, which are either buried or within regions of structural rigidity, will exhibit protection against proteolysis. In contrast, solvent exposed and flexible regions of the protein will be less protected. The assay uses matrix-assisted laser desorption ionization mass spectrometry to map rapidly and accurately the sites of proteolytic cleavage with HPLC. The work has resulted in a host of new findings, culminating in the publication of a full-length report in the journal Protein Science. A computer-designed figure from the paper was accorded the cover of the journal.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-23
Application #
5221593
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

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