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

Research output: Contribution to journalArticle

1 Citation (Scopus)

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

Fingerprint

Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Chromogranins
Proteome
Knockout Mice
Kidney
Imaging techniques
Proteins
Immunohistochemistry
Design of experiments
Nucleic Acids
Carrier Proteins
Genes
Aging of materials
Frozen Sections
Enzymes
Research Design
Western Blotting
Genotype
Mutation
Research

Keywords

  • Kidney
  • Klotho
  • MALDI-IMS

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS. / Fujino, Yoko; Minamizaki, Tomoko; Hayashi, Ikue; Kawakami, Asako; Miyaji, Takaaki; Sakurai, Kaoru; Yoshioka, Hirotaka; Kozai, Katsuyuki; Okada, Mitsugi; Yoshiko, Yuji.

In: Proteomics - Clinical Applications, Vol. 11, No. 7-8, 1600095, 01.07.2017.

Research output: Contribution to journalArticle

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, vol. 11, no. 7-8, 1600095. https://doi.org/10.1002/prca.201600095
Fujino, Yoko ; Minamizaki, Tomoko ; Hayashi, Ikue ; Kawakami, Asako ; Miyaji, Takaaki ; Sakurai, Kaoru ; Yoshioka, Hirotaka ; Kozai, Katsuyuki ; Okada, Mitsugi ; Yoshiko, Yuji. / Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS. In: Proteomics - Clinical Applications. 2017 ; Vol. 11, No. 7-8.
@article{758f1f286d8c45bb99ea39c901526e2f,
title = "Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS",
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.",
keywords = "Kidney, Klotho, MALDI-IMS",
author = "Yoko Fujino and Tomoko Minamizaki and Ikue Hayashi and Asako Kawakami and Takaaki Miyaji and Kaoru Sakurai and Hirotaka Yoshioka and Katsuyuki Kozai and Mitsugi Okada and Yuji Yoshiko",
year = "2017",
month = "7",
day = "1",
doi = "10.1002/prca.201600095",
language = "English",
volume = "11",
journal = "Proteomics - Clinical Applications",
issn = "1862-8346",
publisher = "Wiley-VCH Verlag",
number = "7-8",

}

TY - JOUR

T1 - Comparative proteome analysis of wild-type and klotho-knockout mouse kidneys using a combination of MALDI-IMS and LC-MS/MS

AU - Fujino, Yoko

AU - Minamizaki, Tomoko

AU - Hayashi, Ikue

AU - Kawakami, Asako

AU - Miyaji, Takaaki

AU - Sakurai, Kaoru

AU - Yoshioka, Hirotaka

AU - Kozai, Katsuyuki

AU - Okada, Mitsugi

AU - Yoshiko, Yuji

PY - 2017/7/1

Y1 - 2017/7/1

N2 - 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.

AB - 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.

KW - Kidney

KW - Klotho

KW - MALDI-IMS

UR - http://www.scopus.com/inward/record.url?scp=85017418087&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85017418087&partnerID=8YFLogxK

U2 - 10.1002/prca.201600095

DO - 10.1002/prca.201600095

M3 - Article

VL - 11

JO - Proteomics - Clinical Applications

JF - Proteomics - Clinical Applications

SN - 1862-8346

IS - 7-8

M1 - 1600095

ER -