In vivo pharmaco-proteomic analysis of hydroxyurea induced changes in the sickle red blood cell membrane proteome

Swati S. Ghatpande, Pankaj K. Choudhary, Charles T. Quinn, Steven Goodman

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Hydroxyurea (HU) is an effective drug for the treatment of sickle cell disease (SCD). The main clinical benefit of HU is thought to derive from its capacity to increase fetal hemoglobin (HbF) production. However, other effects leading to clinical benefit, such as improved blood rheology, have been suggested. In order to understand HU-induced changes at the proteomic level, we profiled sickle RBC membranes from of HU-treated and untreated patients. Our previous in vitro profiling studies on sickle RBC membranes identified a significant increase in predominantly anti-oxidant enzymes, protein repair and degradation components and a few RBC cytoskeletal proteins. In the present study, using 2D-DIGE (Two-Dimensional Difference In-Gel Electrophoresis) and tandem mass spectrometry, we detected 32 different proteins that significantly changed in abundance in the HU treatment group. The proteins that significantly increased in abundance were mostly membrane skeletal components involved in the regulation of RBC shape and flexibility, and those showing a significant decrease were components of the protein repair and degradation machinery. RBC palmitoylated membrane protein 55 (p55) is significantly increased in abundance at low (in vitro) and high (in vivo) concentrations of HU. Palmitoylated p55 may be an important target of HU-dependent regulation of the sickle RBC membrane, consistent with our earlier in vitro studies.

Original languageEnglish (US)
Pages (from-to)619-626
Number of pages8
JournalJournal of Proteomics
Volume73
Issue number3
DOIs
StatePublished - Jan 3 2010
Externally publishedYes

Fingerprint

Hydroxyurea
Proteome
Cell membranes
Proteomics
Blood
Erythrocytes
Cell Membrane
Two-Dimensional Difference Gel Electrophoresis
Membranes
Proteins
Proteolysis
Repair
Degradation
Fetal Hemoglobin
Cytoskeletal Proteins
Rheology
Sickle Cell Anemia
Tandem Mass Spectrometry
Electrophoresis
Oxidants

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry

Cite this

In vivo pharmaco-proteomic analysis of hydroxyurea induced changes in the sickle red blood cell membrane proteome. / Ghatpande, Swati S.; Choudhary, Pankaj K.; Quinn, Charles T.; Goodman, Steven.

In: Journal of Proteomics, Vol. 73, No. 3, 03.01.2010, p. 619-626.

Research output: Contribution to journalArticle

Ghatpande, Swati S. ; Choudhary, Pankaj K. ; Quinn, Charles T. ; Goodman, Steven. / In vivo pharmaco-proteomic analysis of hydroxyurea induced changes in the sickle red blood cell membrane proteome. In: Journal of Proteomics. 2010 ; Vol. 73, No. 3. pp. 619-626.
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