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 language | English |
|---|---|
| Article number | 1600095 |
| Journal | Proteomics - Clinical Applications |
| Volume | 11 |
| Issue number | 7-8 |
| DOIs | |
| Publication status | Published - Jul 1 2017 |
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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 journal › Article
}
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
C2 - 28276159
AN - SCOPUS:85017418087
VL - 11
JO - Proteomics - Clinical Applications
JF - Proteomics - Clinical Applications
SN - 1862-8346
IS - 7-8
M1 - 1600095
ER -