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Proteomic Analysis of Differentially Expressed Proteins in Human Lung Cells Following Formaldehyde Treatment 원문보기

Molecular & cellular toxicology, v.3 no.4, 2007년, pp.238 - 245  

Jeon, Yu-Mi (Department of Biotechnology, SoonChunHyang University) ,  Ryu, Jae-Chun (Cellular and Molecular Toxicology Laboratory, Korea Institute of Science and Technology) ,  Lee, Mi-Young (Department of Biotechnology, SoonChunHyang University)

Abstract ▼ AI-Helper 아이콘AI-Helper

Chronic formaldehyde inhalation studies have suggested its relativity to teratogenicity, cancer incidence, neurodegenerative and vascular disorders. Many toxicological data on the formaldehyde toxicity are available, but proteomic results showing complete protein profiles are limited. Therefore, alt...

주제어

#Formaldehyde   #Human lung epithelial cell   #Toxicoproteomics  

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제안 방법

  • The sample solution was applied on immobilized pH 3-10 nonlinear gradient drystrips using an IPG phor system (GE Healthcare Biosciences). Focusing was performed using the following steps: rehydration for 12 hrs, 250 V/100 Vhr, 500 V/500 Vhr, 1,000 V/1,000 Vhr, 8,000 V/38,000 Vhr. After IEF, the individual IPG strips were equilibrated in 6 M urea, 2% SDS, 50 mM Tris-HCl (pH 8.
  • The gels were visualized using CBB-G250 (Coomassie brilliant blue G-250) staining method. The stained gels were scanned using a UMAX scanner (UMAX Technologies, Plano, TX), and the data were analyzed using Image Master 2D Elite software (GE Healthcare Biosciences).

대상 데이터

  • Tryptic peptides derived from protein spots were analyzed and amino acid sequences were deduced using a de novo peptide-sequencing program, PepSeq (Gibbsland). To identify the proteins, sequences were searched against the NCBInr and EST databases using the PROFOUND search program (http://www.rockefeller.edu/labheads/chait/novel_tandem.php) and BLAST.

이론/모형

  • Peptide mass fingerprints and Post-Source Decay (PSD) tryptic peptides were generated by matrix assisted laser desorption/ionization-time-of-flight-mass spectrometry (MA-LDI-TOF-MS) using Ettan MALDI-TOF (GE Healthcare Biosciences). Tryptic peptides derived from protein spots were analyzed and amino acid sequences were deduced using a de novo peptide-sequencing program, PepSeq (Gibbsland).
  • The gels were visualized using CBB-G250 (Coomassie brilliant blue G-250) staining method. The stained gels were scanned using a UMAX scanner (UMAX Technologies, Plano, TX), and the data were analyzed using Image Master 2D Elite software (GE Healthcare Biosciences).
  • To study the differential expression of proteins in lung cell after formaldehyde treatment, proteomic analysis was performed using high-resolution 2-DE. Figure 2 shows 2-DE images of the liver proteins following treatment of 100 µM formaldehyde for 0, 12, 24 and 48 hrs.
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