Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS

Yoko Fujino, Tomoko Minamizaki, Ikue Hayashi, Asako Kawakami, Takaaki Miyaji, Kaoru Sakurai, Hirotaka Yoshioka, Katsuyuki Kozai, Mitsugi Okada, Yuji Yoshiko

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Abstract

Purpose: Mutation of the klotho gene in mice elicits a syndrome resembling accelerated human aging. However, there is limited evidence for the role of Klotho in the kidney. We conducted a comparative proteome analysis of wild-type (WT) and klotho-knockout (kl−/−) mouse kidneys to identify proteins involved in Klotho deficiency. Experimental design: MALDI imaging MS (MALDI-IMS) of frozen kidney sections from 7-wk-old male WT and kl−/− mice was used to determine genotype-specific differences in the MS distribution. Proteins uniquely distributed in kl−/− kidneys were identified by subsequent analysis of adjacent trypsinized sections by MALDI-IMS in combination with LC-MS/MS. Immunohistochemistry and western blotting were adopted in qualitative and quantitation analysis. Results: Ninety-seven and 69 proteins identified by LC-MS/MS were matched to the MALDI-IMS spectra in WT and kl−/− mouse kidneys, respectively. Among protein types matched, nucleic acid binding proteins were most abundant, followed by enzymes. We identified secretogranin-1 (SCG1), which was predominately distributed in the glomeruli and renal tubules of kl−/− mouse kidneys. Immunohistochemistry for SCG1 mirrored images of MALDI-IMS. Conclusions: SCG1 may be a candidate protein involved in Klotho deficiency. Although further research is needed to investigate the role of SCG1 in the kidney, we show the usefulness of MALDI-IMS combined with LC-MS/MS.

Original languageEnglish
Article number1600095
JournalProteomics - Clinical Applications
Volume11
Issue number7-8
DOIs
Publication statusPublished - Jul 1 2017

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Keywords

  • Kidney
  • Klotho
  • MALDI-IMS

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Fujino, Y., Minamizaki, T., Hayashi, I., Kawakami, A., Miyaji, T., Sakurai, K., Yoshioka, H., Kozai, K., Okada, M., & Yoshiko, Y. (2017). Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS. Proteomics - Clinical Applications, 11(7-8), [1600095]. https://doi.org/10.1002/prca.201600095