Method Development of Efficient Protein Extraction in Bone Tissue for Proteome Analysis
Abstract
Exploring bone proteome is an important and challenging task for understanding the mechanisms of physiological/pathological process of bone tissue. However, classical methods of protein extraction for soft tissues and cells are not applicable for bone tissue. Therefore, method development of efficient protein extraction is critical for bone proteome analysis. We found in this study that the protein extraction efficiency was improved significantly when bone tissue was demineralized by hydrochloric acid (HCl). A sequential protein extraction method was developed for large-scale proteome analysis of bone tissue. The bone tissue was first demineralized by HCl solution and then extracted using three different lysis buffers. As large amounts of acid soluble proteins also presented in the HCl solution, besides collection of proteins in the extracted lysis buffers, the proteins in the demineralized HCl solution were also collected for proteome analysis. Automated 2D-LC-MS/MS analysis of the collected protein fractions resulted in the identification of 6202 unique peptides which matched 2479 unique proteins. The identified proteins revealed a broad diversity in the protein identity and function. More than 40 bone-specific proteins and 15 potential protein biomarkers previously reported were observed in this study. It was demonstrated that the developed extraction method of proteins in bone tissue, which was also the first large-scale proteomic study of bone, was very efficient for comprehensive analysis of bone proteome and might be helpful for clarifying the mechanisms of bone diseases.
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Proteomic analysis identified more than 100 organic ECM proteins, with collagen type I and noncollagenous proteins being the major constituents.
“proteomic analysis have led to the identification of more than 100 organic ECM proteins,197 with collagen type I (Coll-1) and noncollagenous proteins (NCPs) being the major constituents.”
From Bone remodeling: an operational process ensuring survival and bone mechanical competence (2022) · cites this paper as [197]
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- Authors
- Jiang, X., Ye, M., Jiang, X., Liu, G., Feng, S., Cui, L., & Zou, H.
- Affiliations
- Dalian Institute of Chemical Physics (CN)
- Corresponding author
- Zou, Hanfa
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2007-05-08
- Volume
- 6
- Issue
- 6
- Pages
- 2287-2294
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- OpenAlex
- W2126829660
- Cited by
- 144
- References
- 53
- Citations by year
- 2026: 5 · 2025: 4 · 2024: 9 · 2023: 10 · 2022: 5 · 2021: 8 · 2020: 5 · 2019: 21 · 2018: 13 · 2017: 7 · 2016: 11 · 2015: 7 · 2014: 8 · 2013: 7 · 2012: 6
- FWCI
- 2.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Proteome Research (publishedVersion)
- PubMed (publishedVersion)
- Dalian Institute of Chemical Physics, CAS Institutional Repository (submittedVersion)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Health Sciences › Medicine › Rheumatology › Bone and Dental Protein Studies
- Health Sciences › Medicine › Orthopedics and Sports Medicine › Bone health and osteoporosis research
- Keywords
- bone proteome, protein extraction, sequential extraction, acid-soluble proteins, bone-specific protein, bone disease, 2D-LC-MS/MS
- MeSH terms
- Animals, Bone and Bones, Dogs, Proteins, Proteome, Proteomics
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