Developmental regulation of FKBP65. An ER-localized extracellular matrix binding-protein.
FKBP65 (65-kDa FK506-binding protein) is a member of the highly conserved family of intracellular receptors called immunophilins. All have the property of peptidyl-prolyl cis-trans isomerization, and most have been implicated in folding and trafficking events. In an earlier study, we identified that FKBP65 associates with the extracellular matrix protein tropoelastin during its transport through the cell. In the present study, we have carried out a detailed investigation of the subcellular localization of FKBP65 and its relationship to tropoelastin. Using subcellular fractionation, Triton X-114 phase separation, protease protection assays, and immunofluorescence microscopy (IF), we have identified that FKBP65 is contained within the lumen of the endoplasmic reticulum (ER). Subsequent IF studies colocalized FKBP65 with tropoelastin and showed that the two proteins dissociate before reaching the Golgi apparatus. Immunohistochemical localization of FKBP65 in developing lung showed strong staining of vascular and airway smooth muscle cells. Similar areas stained positive for the presence of elastic fibers in the extracellular matrix. The expression of FKBP65 was investigated during development as tropoelastin is not expressed in adult tissues. Tissue-specific expression of FKBP65 was observed in 12-d old mouse tissues; however, the pattern of expression of FKBP65 was not restricted to those tissues expressing tropoelastin. This suggests that additional ligands for FKBP65 likely exist within the ER. Remarkably, in the adult tissues examined, FKBP65 expression was absent or barely detectable. Taken together, these results support an ER-localized FKBP65-tropoelastin interaction that occurs specifically during growth and development of tissues.
Patterson
C E
CE
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
Schaub
T
T
Coleman
E J
EJ
Davis
E C
EC
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006056:Golgi Apparatus / Q000378:metabolism; D007425:Intracellular Membranes; D008168:Lung / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000378:metabolism; D019696:Peptidylprolyl Isomerase; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism*; D014333:Tropoelastin / Q000378:metabolism
D014333:Tropoelastin; D022021:Tacrolimus Binding Proteins; C491675:Fkbp10 protein, mouse; D019696:Peptidylprolyl Isomerase