Identification and characterization of rodent ABCA1 in isolated type II pneumocytes

Bortnick, Anna E.; Favari, Elda; Jiang, Tao; Francone, Omar L.; Reilly, Muredach P; Zhang, Yuzhen; Rothblat, George H.; Bates, Sandra R.

doi:10.1152/ajplung.00077.2003

Cited by 57 publications

(58 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mouse AEC I and AEC II were isolated from male C57BL/6 mice (6-8 weeks of age) according to the previously reported procedure, with some modification (Bortnick et al, 2003). Mice were anesthetized with ketamine (80 mg/kg of body weight) and xylazine (10 mg/kg of body weight).…”

Section: Isolation Of Mouse Aec I and Aec Iimentioning

confidence: 99%

Purinergic P2X7 receptor regulates lung surfactant secretion in a paracrine manner

Mishra

Chintagari

Guo

et al. 2011

Journal of Cell Science

View full text Add to dashboard Cite

SummaryAlveolar epithelium is composed of alveolar epithelial cells of type I (AEC I) and type II (AEC II). AEC II secrete lung surfactant by means of exocytosis. P2X 7 receptor (P2X 7 R), a P2 purinergic receptor, has been implicated in the regulation of synaptic transmission and inflammation. Here, we report that P2X 7 R, which is expressed in AEC I but not AEC II, is a novel mediator for the paracrine regulation of surfactant secretion in AEC II. In primary co-cultures of AEC I and AEC II benzoyl ATP (BzATP; an agonist of P2X 7 R) increased surfactant secretion, which was blocked by the P2X 7 R antagonist Brilliant Blue G. This effect was observed in AEC II cocultured with human embryonic kidney HEK-293 cells stably expressing rat P2X 7 R, but not when co-cultured with AEC I in which P2X 7 R was knocked down or in co-cultures of AEC I and AEC II isolated from P2X 7 R -/-mice. BzATP-mediated secretion involved P2Y 2 receptor signaling because it was reduced by the addition of the ATP scavengers apyrase and adenosine deaminase and the P2Y 2 receptor antagonist suramin. However, the stimulation with BzATP might also release other substances that potentially increase surfactant secretion as a greater stimulation of secretion was observed in AEC II incubated with BzATP when co-cultured with E10 or HEK-293-P2X 7 R cells than with ATP alone. P2X 7 R -/-mice failed to increase surfactant secretion in response to hyperventilation, pointing to the physiological relevance of P2X 7 R in maintaining surfactant homeostasis in the lung. These results suggest that the activation of P2X 7 R increases surfactant secretion by releasing ATP from AEC I and subsequently stimulating P2Y 2 receptors in AEC II.

show abstract

Section: Isolation Of Mouse Aec I and Aec Iimentioning

confidence: 99%

Purinergic P2X7 receptor regulates lung surfactant secretion in a paracrine manner

Mishra

Chintagari

Guo

et al. 2011

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…Mouse Type 2 Cells-Primary mouse type 2 pneumocytes, isolated from C57BL/6 mouse lungs using dispase, mesh filtration, and panning over IgG-coated and uncoated plates as described previously (29,30), were kindly provided by Dr. Sandra Bates and the Cell Culture Core at the Institute for Environmental Medicine at the University of Pennsylvania. Cell pellets were prepared for Western blotting from freshly isolated cells by detergent lysis as previously published (2,8).…”

Section: Cell Models and Transfectionmentioning

confidence: 99%

Anterograde Transport of Surfactant Protein C Proprotein to Distal Processing Compartments Requires PPDY-mediated Association with Nedd4 Ubiquitin Ligases

Kotorashvili

Russo

Mulugeta

et al. 2009

Journal of Biological Chemistry

View full text Add to dashboard Cite

Biosynthesis of surfactant protein C (SP-C) by alveolar type 2 cells requires proteolytic processing of a 21-kDa propeptide (proSP-C 21 ) in post-Golgi compartments to yield a 3.7-kDa mature form. Scanning alanine mutagenesis, binding assays, and co-immunoprecipitation were used to characterize the proSP-C targeting domain. Delivery of proSP-C 21 to distal processing organelles is dependent upon the NH 2 -terminal cytoplasmic SP-C propeptide, which contains a conserved PPDY motif. In A549 cells, transfection of EGFP/proSP-C 21 constructs containing polyalanine substitution for Glu 11 -Thr 18 , 13 PPDY 16 , or 14 P, 16 Y produced endoplasmic reticulum retention of the fusion proteins. Protein-protein interactions of proSP-C with known WW domains were screened using a solid-phase array that revealed binding of the proSP-C NH 2 terminus to several WW domains found in the Nedd4 family of E3 ligases. Specificity of the interaction was confirmed by co-immunoprecipitation of proSP-C and Nedd4 or Nedd4-2 in epithelial cell lines. By Western blotting and reverse transcription-PCR, both forms were detected in primary human type 2 cells. Knockdown of Nedd4-2 by small interference RNA transfection of cultured human type 2 cells blocked processing of 35 S-labeled proSP-C 21. Mutagenesis of potential acceptor sites for ubiquitination in the cytosolic domain of proSP-C (Lys 6 , Lys 34 , or both) failed to inhibit trafficking of EGFP/proSP-C 21 . These results indicate that PPDYmediated interaction with Nedd4 E3-ligases is required for trafficking of proSP-C. We speculate that the Nedd4/proSP-C tandem is part of a larger protein complex containing a ubiquitinated component that further directs its transport. Surfactant protein C (SP-C)3 is a hydrophobic lung-specific protein that co-isolates with the biophysically active phospholipid fraction of pulmonary surfactant (1). The 3.7-kDa alveolar form of SP-C (SP-C 3.7 or mature SP-C) is a 35-amino acid, valine-rich peptide that results from synthesis of a 197 amino acid precursor form (proSP-C 21 ). Within the endoplasmic reticulum (ER), proSP-C 21 exists as a type II bitopic transmembrane protein (N cyt /C lumen ), which remains integrally membrane-associated while it is extensively processed by four post-translational endoproteolytic cleavages that take place in post-Golgi compartments (2-6). The resulting mature protein is transferred into the lumen of lamellar bodies for secretion into the alveolar space together with surfactant phospholipid and SP-B (7). Thus, the complete biosynthesis of SP-C 3.7 is critically dependent upon oligomeric sorting and targeting of proSP-C 21 to subcellular processing compartments distal to the ER/Golgi.Using deletional mutagenesis, we had previously shown that the NH 2 -terminal propeptide flanking domain (NTP) of proSP-C, which is oriented into the cytosol, contained potential targeting motifs for post-Golgi trafficking (8, 9). The candidate region included the amino acid sequence Met 10 -Thr 18 and, when present, was sufficient to direct transf...

show abstract

“…Alveolar type 2 epithelial cells were isolated from mouse lungs by a previously described procedure using endotracheal dispase for lung digestion (16). Cell isolates were seeded onto type I collagen-coated 35 mm dishes in Ham's F-12 culture medium supplemented with 15 mM HEPES, 0.8 mM CaCl 2 , 0.25% BSA, 5 g/ml insulin, 5 g/ml transferrin, 5 ng/ml sodium selenite, and 2% mouse serum and incubated for 24 h before study.…”

Section: Animals and Cell Isolationmentioning

confidence: 99%

Altered lung phospholipid metabolism in mice with targeted deletion of lysosomal-type phospholipase A2

Fisher

Dodia

Feinstein

et al. 2005

Journal of Lipid Research

107

View full text Add to dashboard Cite

Identification and characterization of rodent ABCA1 in isolated type II pneumocytes

Cited by 57 publications

References 55 publications

Purinergic P2X7 receptor regulates lung surfactant secretion in a paracrine manner

Purinergic P2X7 receptor regulates lung surfactant secretion in a paracrine manner

Anterograde Transport of Surfactant Protein C Proprotein to Distal Processing Compartments Requires PPDY-mediated Association with Nedd4 Ubiquitin Ligases

Altered lung phospholipid metabolism in mice with targeted deletion of lysosomal-type phospholipase A2

Contact Info

Product

Resources

About