title: Effect of FKBP65, a putative elastin chaperone, on the coacervation of tropoelastin in vitro.
abstract: FKBP65 is a protein of the endoplasmic reticulum that is relatively abundant in elastin-producing cells and is associated with tropoelastin in the secretory pathway. To test an earlier suggestion by Davis and co-workers that FKBP65 could act as an intracellular chaperone for elastin, we obtained recombinant FKBP65 (rFKBP65) by expressing it in E. coli and examined its effect on the coacervation characteristics of chicken aorta tropoelastin (TE) using an in vitro turbidimetric assay. Our results reveal that rFKBP65 markedly promotes the initiation of coacervation of TE without significantly affecting the temperature of onset of coacervation. This effect shows saturation at a 1:2 molar ratio of TE to rFKBP65. By contrast, FKBP12, a peptidyl prolyl isomerase, has a negligible effect on TE coacervation. Moreover, the effect of rFKBP65 on TE coacervation is unaffected by the addition of rapamycin, an inhibitor of peptidyl prolyl isomerase (PPIase) activity. These observations rule out the involvement of the PPIase activity of rFKBP65 in modulating the coacervation of TE. Additional experiments using a polypeptide model of TE showed that rFKBP65, while promoting coacervation, may retard the maturation of this model polypeptide into larger aggregates. Based on these results, we suggest that FKBP65 may act as an elastin chaperone in vivo by controlling both the coacervation and the maturation stages of its self-assembly into fibrils.
authors.affiliation: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada.
authors.forename: Kevin L Y
authors.initials: KL
authors.lastname: Cheung
authors.forename: Matthew
authors.initials: M
authors.lastname: Bates
authors.forename: Vettai S
authors.initials: VS
authors.lastname: Ananthanarayanan
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism; D002645:Chickens; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004549:Elastin / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019696:Peptidylprolyl Isomerase / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012155:Reticulin / Q000737:chemistry*; D055571:Secretory Pathway / Q000187:drug effects; D020123:Sirolimus / Q000494:pharmacology; D022021:Tacrolimus Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014333:Tropoelastin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D018832:Molecular Chaperones; D011994:Recombinant Proteins; D012155:Reticulin; D014333:Tropoelastin; D004549:Elastin; D022021:Tacrolimus Binding Proteins; C491675:Fkbp10 protein, mouse; D019696:Peptidylprolyl Isomerase; D020123:Sirolimus