Collect. Czech. Chem. Commun. 2001, 66, 1720-1728

Identification of HL60 Proteins Affected by 5-Aminolevulinic Acid-Based Photodynamic Therapy Using Mass Spectrometric Approach

Petr Haladaa, Petr Mana, Dana Grebeňováb, Zbyněk Hrkalb and Vladimír Havlíčeka,*

a Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic
b Department of Cellular Biochemistry, Institute of Hematology and Blood Transfusion, U Nemocnice 1, CZ-128 20 Prague 2, Czech Republic


A combination of mass spectrometric techniques was used for identification of HL60 leukemia cell proteins affected by 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT). We compared two-dimensional electrophoresis (2-DE) protein maps of ALA-treated non-irradiated and irradiated cells and found extensive changes in the proteome of HL60 cells. The silver-stained 2-DE pattern of HL60 proteins contained more than 1 350 spots. Matrix-assisted laser desorption/ionisation mass spectrometry and microcapillary liquid chromatography/tandem mass spectrometry have identified twelve proteins differing in their intensity or position following ALA-PDT. Several endoplasmic reticulum, mitochondrial, ribosomal and cytoplasmic proteins were determined showing the impact of ALA-PDT-mediated cytotoxicity on some cellular pathways.

Keywords: Mass spectrometry; Matrix-assisted laser desorption/ionisation; MALDI; Electrospray ionisation; Protein identification; Peptide mass fingerprinting; Photodynamic therapy.

References: 32 live references.