Adenylate cyclase stimulating activity immunologically similar to parathyroid hormone-related peptide can be extracted from fetal rat long bones

Autor: Jacques Corvilain, Rafik Karmali, Thierry Pepersack, Pierre Bergmann, Nicole Nijs-de Wolf
Rok vydání: 2009
Předmět:
Ulna -- embryology
medicine.medical_specialty
Kidney Cortex
Endocrinology
Diabetes and Metabolism

Proteolysis
Blotting
Western

Proteins -- isolation & purification
Bone Matrix
Adenylate kinase
Parathyroid hormone
Ulna
Biology
Cyclase
Bone and Bones
Bone resorption
Bone Matrix -- chemistry
Ostéologie
Internal medicine
Cyclic AMP
medicine
Animals
Orthopedics and Sports Medicine
Proteins -- immunology
Imagerie médicale
radiologie
tomographie

Chromatography
High Pressure Liquid

Antiserum
Analysis of Variance
medicine.diagnostic_test
Parathyroid hormone-related protein
Immune Sera
Cell Membrane
Bone and Bones -- embryology
Parathyroid Hormone-Related Protein
Proteins -- pharmacology
Proteins
Rats
Inbred Strains

Cyclic AMP -- biosynthesis
Rats
Resorption
Bone and Bones -- chemistry
Endocrinology
Adenylate Cyclase -- metabolism
Electrophoresis
Polyacrylamide Gel

Kidney Cortex -- enzymology
Cell Membrane -- enzymology
Adenylyl Cyclases
Zdroj: Journal of Bone and Mineral Research, 6 (9
ISSN: 0884-0431
DOI: 10.1002/jbmr.5650060905
Popis: We observed that culture medium conditioned with fetal rat long bones contained peptides immunologically related to the parathyroid hormone-related peptide of malignancy (PTHrP) and stimulated cyclic AMP production in canine renal cortical membranes. Because the adenylate cyclase stimulating activity (CSA) of the medium increased when bone resorption was stimulated, it was suspected that these peptides were stored in the matrix and released during the resorption process. In this work, we extracted the noncollagenous proteins of fetal rat long bones and found that the extract contained significant amounts of CSA. The biologic activity of the extract was abolished after trypsin digestion and eluted at 24 and 37 kD on filtration HPLC. The CSA of bone extract and of both HPLC peaks could be inhibited by 3-34 and 7-34 parathyroid hormone analogs. It was not blocked by an antiserum directed against the N-terminal region of parathyroid hormone, but it was significantly inhibited after an overnight preincubation with an antiserum directed against the 1-11 fragment of PTHrP. One band migrating at 18 kD could be visualized after SDS-PAGE electrophoresis of bone extract and immunoblotting with the anti-PTHrP antiserum. We conclude that an adenylate cyclase stimulator immunologically similar to PTHrP is present in the matrix of fetal rat long bones. Adenylate cyclase stimulating peptides of lower molecular weight found in bone-conditioned medium could be active fragments formed by proteolysis during the resorption process.
Journal Article
Research Support, Non-U.S. Gov't
FLWNA
info:eu-repo/semantics/published
Databáze: OpenAIRE