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Naina Adren Pinto(진단검사의학교실) Oncotarget
등록일 : 2017-07-17 오후 1:38:00       조회 : 20
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제1저자 Naina Adren Pinto(진단검사의학교실, BK21)
교신저자 용동은(진단검사의학교실)

Oncotarget. 2017 Jul 5. doi: 10.18632/oncotarget.19005. [Epub ahead of print]
Whole genome and transcriptome analysis reveal MALDI-TOF MS and SDS-PAGE have limited performance for the detection of the key outer membrane protein in carbapenem-resistant Klebsiella pneumoniae isolates.
Pinto NA1,2, D''Souza R1, Hwang IS1,2, Choi J1, In YH3, Park HS3, Ryu CM4,5, Yong D1, Lee K1.

Author information
1Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea.2Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul, Korea.3ASTA Corporation, Suwon, Korea.4Molecular Phytobacteriology Laboratory, KRIBB, Daejeon, Korea.5Biosystems and Bioengineering Program, School of Science, University of Science and Technology, Daejeon, Korea.

To detect the outer membrane protein (OMP), which plays a key role in carbapenem resistance, whole-genome and transcriptome analysis of the clinical carbapenem-resistant Klebsiella pneumoniae was carried out. The index strain lacked both OmpK35 and OmpK36, whereas the other strains lacked only OmpK35. After SDS-PAGE, the putative OMP bands were excised and identified as OmpA and OmpK36. MALDI-TOF MS showed peaks at ~36 and ~38 kDa that corresponded to OmpA and OmpK36, respectively. In all the strains except YMC2014/03/P345, the ~38 kDa peaks were present. The K. pneumoniae ATCC 13883 isolate showed three bands on SDS-PAGE and three corresponding peaks on MALDI-TOF MS. The additional third peak at ~37 kDa corresponding to OmpK35 was observed. To verify OmpK35 peak detection in other K. pneumoniae isolates by MALDI-TOF MS, we analyzed six strains from our laboratory''s strain bank. Whole genome sequence indicated that only two isolates had intact OmpK35. Both MALDI-TOF MS and SDS-PAGE did not show a ~37 kDa peak or an OmpK35 band as observed in the K. pneumoniae ATCC 13883 isolate. Separation using SDS-PAGE showed a single peak representing OmpA. Therefore, both SDS-PAGE and MALDI-TOF MS were not completely reliable for OMP detection because they fail to detect OmpK35. To the best of our knowledge, this is the first report on the performance of SDS-PAGE and MALDI-TOF MS for the detection of OMP''s using whole-genome and RNA sequencing analyses.
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정승수(생리학교실) Cell Rep 2017-07-10