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Updated: Apr 20, 2026
Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis
Published on: July 23, 2015
Proteomic analysis of the extracellular matrix in idiopathic pes equinovarus
Martin Ošt'ádal1, Adam Eckhardt, Jan Herget
1Department of Orthopaedics, 1st Faculty of Medicine, University Hospital Bulovka, Charles University, Budínova 2, 180 00, Prague 8, Czech Republic, martinostadal@yahoo.com.
Insights
Idiopathic pes equinovarus, or clubfoot, involves altered extracellular matrix proteins. This study identified 19 proteins, including collagens I, III, V, VI, and XII, offering insights into clubfoot pathogenesis.
Area of Science:
- Orthopedics
- Biochemistry
- Developmental Biology
Background:
- Idiopathic pes equinovarus (clubfoot) is a congenital foot deformity with unclear pathogenesis.
- Fibroblasts and growth factors are implicated in clubfoot development.
Purpose of the Study:
- To directly analyze the protein composition of the extracellular matrix in contracted clubfoot tissues.
- To identify novel proteins contributing to clubfoot pathology.
Main Methods:
- Tissue samples from 13 infants with idiopathic clubfoot undergoing surgery were analyzed.
- Protein extraction involved digestion and delipidation, followed by chemiluminescent assay for collagen detection.
- Amino acid analysis and identification of 19 extracellular matrix proteins were performed.
Main Results:
- Amino acid analysis indicated a predominance of collagen types I, III, and VI.
- A total of 19 extracellular matrix proteins were identified in clubfoot tissues.
- Key findings include the presence of collagens V, VI, XII, and transforming growth factor β.
Conclusions:
- The extracellular matrix in clubfoot is characterized by a unique protein composition, including several collagen types and growth factors.
- Understanding this protein profile may elucidate clubfoot pathogenesis.
- This research could lead to improved therapeutic strategies for clubfoot.
Abstract:
Idiopathic pes equinovarus is a congenital deformity of the foot and lower leg defined as a fixation of the foot in adduction, supination, and varus. Although the pathogenesis of clubfoot remains unclear, it has been suggested that fibroblasts and growth factors are involved. To directly analyze the protein composition of the extracellular matrix in contracted tissue of patients with clubfoot. A total of 13 infants with idiopathic clubfoot treated with the Ponseti method were included in the present study. Tissue samples were obtained from patients undergoing surgery for relapsed clubfeet. Contracted tissues were obtained from the medial aspect of the talonavicular joint. Protein was extracted after digestion and delipidation using zip-tip C18. Individual collagenous fractions were detected using a chemiluminescent assay. Amino acid analysis of tissue samples revealed a predominance of collagens, namely collagen types I, III, and VI. The high content of glycine and h-proline suggests a predominance of collagens I and III. A total of 19 extracellular matrix proteins were identified. The major result of the present study was the observation that the extracellular matrix in clubfoot is composed of an additional 16 proteins, including collagens V, VI, and XII, as well as the previously described collagen types I and III and transforming growth factor β. The characterization of the general protein composition of the extracellular matrix in various regions of clubfoot may help in understanding the pathogenesis of this anomaly and, thus, contribute to the development of more efficacious therapeutic approaches.
