Antibody raised to AKAAAKAAAKA sequence on tropoelastin recognizes tropoelastin but not mature crosslinked elastin: A new tool in metabolic and structural studies of elastogenesis

Citation
B. Starcher et al., Antibody raised to AKAAAKAAAKA sequence on tropoelastin recognizes tropoelastin but not mature crosslinked elastin: A new tool in metabolic and structural studies of elastogenesis, CONNECT TIS, 40(4), 1999, pp. 273
Citations number
23
Categorie Soggetti
da verificare
Journal title
CONNECTIVE TISSUE RESEARCH
ISSN journal
03008207 → ACNP
Volume
40
Issue
4
Year of publication
1999
Database
ISI
SICI code
0300-8207(1999)40:4<273:ARTASO>2.0.ZU;2-0
Abstract
Tropoelastin, which is secreted from the cell in a soluble form, contains s pecific alanine rich repeat domains that are destined to form covalent desm osine and isodesmosine crosslinks in mature insoluble elastin. We raised a monospecific polyclonal antibody to a AKAAAKAAAKA synthetic peptide (AKA) w hich represents this alanine rich region of tropoelastin. The antibody was reactive with the original peptide antigen and purified tropoelastin, but n ot with mature crosslinked elastin isolated from several animal species. Co nditioned media from chick aorta smooth muscle cells in culture reacted in an ELISA with the AKA antibody, but only in the presence of BAPN to block t he conversion of the epsilon-amino groups to aldehydes, Immunofluorescence demonstrated that the AKA antibody decorated newly deposited tropoelastin a ssembled in fine fibrils in matrix produced by cultured human skin fibrobla sts, EM-immunogold specifically localized this antibody to the immature ela stic fibers present in fetal sheep ductus arteriosus. Moreover, immunohisto chemistry demonstrated that the antibody recognized nonpolymerized tropoela stin assembled on the periphery of elastic fibers in the aorta of chicks ra ised on copper deficient and BAPN containing diets. These studies demonstra te that this new anti-tropoelastin antibody can be used as a useful tool to investigate elastin metabolism where it is important to distinguish betwee n tropoelastin and mature crosslinked elastin.