Role of P63 (CKAP4) in binding of surfactant protein-A to type II pneumocytes.
We have recently described a putative receptor for lung surfactant protein-A (SP-A) on rat type II pneumocytes. The receptor, P63, is a 63-kDa type II transmembrane protein. Coincubation of type II cells with P63 antibody (Ab) reversed the inhibitory effect of SP-A on secretagogue-stimulated surfactant secretion from type II cells. To further characterize SP-A interactions with P63, we expressed recombinant P63 protein in Escherichia coli and generated antibodies to P63. Immunogold electron microscopy confirmed endoplasmic reticulum and plasma membrane localization of P63 in type II cells with prominent labeling of microvilli. Binding characteristics of iodinated SP-A to type II cells in the presence of P63 Ab were determined. Binding (4 degrees C, 1 h) of (125)I-SP-A to type II cells demonstrated both specific (calcium-dependent) and nonspecific (calcium-independent) components. Ab to P63 protein blocked the specific binding of (125)I-SP-A to type II cells and did not change the nonspecific SP-A association. A549 cells, a pneumocyte model cell line, expressed substantial levels of P63 and demonstrated specific binding of (125)I-SP-A that was inhibited by the P63 Ab. The secretagogue (cAMP)-stimulated increase in calcium-dependent binding of SP-A to type II cells was blocked by the presence of P63 Ab. Transfection of type II cells with small interfering RNA to P63 reduced P63 protein expression, attenuated P63-specific SP-A binding, and reversed the ability of SP-A to prevent surfactant secretion from the cells. Our results further substantiate the role of P63 as an SP-A receptor protein localized on the surface of lung type II cells.
Bates
Sandra R
SR
Institute for Environmental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. batekenn@mail.med.upenn.edu
Kazi
Altaf S
AS
Tao
Jian-Qin
JQ
Yu
Kevin J
KJ
Gonder
Daniel S
DS
Feinstein
Sheldon I
SI
Fisher
Aron B
AB
D000230:Adenocarcinoma; D000818:Animals; D045744:Cell Line, Tumor; D006801:Humans; D008168:Lung / Q000502:physiology*; D008175:Lung Neoplasms; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D018613:Microscopy, Confocal; D016253:Microscopy, Immunoelectron; D010957:Plasmids; D037662:Pulmonary Surfactant-Associated Protein A / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism
C081876:CKAP4 protein, human; D008565:Membrane Proteins; D037662:Pulmonary Surfactant-Associated Protein A; D034741:RNA, Small Interfering; D011994:Recombinant Proteins