Isolation of human type X collagen and immunolocalization in fetal human cartilage

Thorsten KIRSCH; Klaus VON DER MARK

doi:10.1111/j.1432-1033.1991.tb15852.x

Isolation of human type X collagen and immunolocalization in fetal human cartilage

Thorsten KIRSCH, Klaus VON DER MARK

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

Abstract

Type X Collagen was extracted with 1 M NaCl and 10 mM dithiothreitol at neutral pH from fetal human growth plate cartilage and purified to homogeneity by gel filtration and anion‐exchange chromatography. The purified protein migrates in SDS/polyacrylamide gels with an apparent M_r of 66000 under reducing conditions, and as a high‐M_r oligomer under non‐reducing conditions. Purified collagenase digests most of the molecule; pepsin digestion at 4°C decreases the M_r of the monomer to 53000. A rabbit antiserum was raised against purified human type X collagen; the IgG fraction was specific for this collagen by criteria of ELISA and immunoblotting after absorption with collagen types I, II, VI, IX and XI. Immunohistological studies localized type X collagen exclusively in the zone of hypertrophic and calcifying cartilage.

Original language	English (US)
Pages (from-to)	575-580
Number of pages	6
Journal	European Journal of Biochemistry
Volume	196
Issue number	3
DOIs	https://doi.org/10.1111/j.1432-1033.1991.tb15852.x
State	Published - Mar 1991

ASJC Scopus subject areas

Biochemistry

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10.1111/j.1432-1033.1991.tb15852.x

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abstract = "Type X Collagen was extracted with 1 M NaCl and 10 mM dithiothreitol at neutral pH from fetal human growth plate cartilage and purified to homogeneity by gel filtration and anion‐exchange chromatography. The purified protein migrates in SDS/polyacrylamide gels with an apparent Mr of 66000 under reducing conditions, and as a high‐Mr oligomer under non‐reducing conditions. Purified collagenase digests most of the molecule; pepsin digestion at 4°C decreases the Mr of the monomer to 53000. A rabbit antiserum was raised against purified human type X collagen; the IgG fraction was specific for this collagen by criteria of ELISA and immunoblotting after absorption with collagen types I, II, VI, IX and XI. Immunohistological studies localized type X collagen exclusively in the zone of hypertrophic and calcifying cartilage.",

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AB - Type X Collagen was extracted with 1 M NaCl and 10 mM dithiothreitol at neutral pH from fetal human growth plate cartilage and purified to homogeneity by gel filtration and anion‐exchange chromatography. The purified protein migrates in SDS/polyacrylamide gels with an apparent Mr of 66000 under reducing conditions, and as a high‐Mr oligomer under non‐reducing conditions. Purified collagenase digests most of the molecule; pepsin digestion at 4°C decreases the Mr of the monomer to 53000. A rabbit antiserum was raised against purified human type X collagen; the IgG fraction was specific for this collagen by criteria of ELISA and immunoblotting after absorption with collagen types I, II, VI, IX and XI. Immunohistological studies localized type X collagen exclusively in the zone of hypertrophic and calcifying cartilage.

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Isolation of human type X collagen and immunolocalization in fetal human cartilage

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