Immunoferritin binding to proteoglycan monomers. An electron microscopic study

Joseph A Buckwalter; A Robin Poole; Agnes Reiner; Lawrence C Rosenberg

doi:10.1016/S0021-9258(18)33848-1

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Immunoferritin binding to proteoglycan monomers. An electron microscopic study

Journal article

Open access

Peer reviewed

Immunoferritin binding to proteoglycan monomers. An electron microscopic study

Joseph A Buckwalter, A Robin Poole, Agnes Reiner and Lawrence C Rosenberg

The Journal of biological chemistry, Vol.257(18), pp.10529-10532

09/25/1982

DOI: 10.1016/S0021-9258(18)33848-1

PMID: 7107621

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Abstract

Antibody to native bovine nasal cartilage proteoglycan monomer was shown by enzyme-linked immunosorbent assay to react with the purified hyaluronic acid binding region of the monomer. Antibody was digested with pepsin to produce F(ab')2 and labeled with glutaraldehyde-activated ferritin. F(ab')2 and F(ab')2-ferritin were reduced and alkylated to render them monovalent (Fab'). Antibody Fab' binding to native proteoglycan monomer was studied by electron microscopy of monomer reacted with ferritin-labeled antibody Fab' spread in a cytochrome c film. Ferritin-labeled antibody Fab' bound primarily at one end of the proteoglycan monomer. This binding was partly inhibitable by unlabeled monovalent antibody Fab', demonstrating immunospecificity. The end of the monomer with bound ferritin sometimes appeared as the thin segment, previously observed to bind to hyaluronic acid. These observations indicate that the hyaluronic acid binding region is at only one end of each proteoglycan monomer and that ferritin-labeled antibody Fab' selectively attaches to this part of native proteoglycan monomers. This methodology should be useful for future structural studies of isolated proteoglycans.

Cartilage - analysis

Animals

Enzyme-Linked Immunosorbent Assay

Immune Sera

Cattle

Immunoglobulin G

Nose

Proteoglycans - analysis

Immunoglobulin Fab Fragments

Microscopy, Electron

Ferritins

Details

Title: Subtitle: Immunoferritin binding to proteoglycan monomers. An electron microscopic study
Creators: Joseph A Buckwalter
A Robin Poole
Agnes Reiner
Lawrence C Rosenberg
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.257(18), pp.10529-10532
DOI: 10.1016/S0021-9258(18)33848-1
PMID: 7107621
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: CA23945 / NCI NIH HHS AM21498 / NIADDK NIH HHS HL16387 / NHLBI NIH HHS
Language: English
Date published: 09/25/1982
Academic Unit: Stead Family Department of Pediatrics; Orthopedics and Rehabilitation; Injury Prevention Research Center
Record Identifier: 9984040479502771

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