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J. Biol. Chem., Vol. 280, Issue 41, 34447-34457, October 14, 2005

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Identification of the Surfactant Protein A Receptor 210 as the Unconventional Myosin 18A^*

Ching-Hui Yang, Jacek Szeliga, Jeremy Jordan, Shawn Faske, Zvjezdana Sever-Chroneos, Bre Dorsett, Robert E. Christian¶, Robert E. Settlage¶, Jeffrey Shabanowitz¶, Donald F. Hunt||, Jeffrey A. Whitsett, and Zissis C. Chroneos1

From the Center of Biomedical Research, University of Texas Health Center, Tyler, Texas 75708-3154, Section of Neonatology, Perinatal and Pulmonary Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229, the ^¶Chemistry Department and the ^||Departments of Chemistry and Pathology, University of Virginia, Charlottesville, Virginia 22904

Mass spectrometric characterization of the surfactant protein A (SP-A) receptor 210 (SP-R210) led to the identification of myosin (Myo) XVIIIA and nonmuscle myosin IIA. Antibodies generated against the unique C-terminal tail of MyoXVIIIA revealed that MyoXVIIIA, MyoIIA, and SP-R210 have overlapping tissue distribution, all being highly expressed in myeloid cells, bone marrow, spleen, lymph nodes, and lung. Western blot analysis of COS-1 cells stably transfected with either MyoXVIIIA or MyoIIA indicated that SP-R210 antibodies recognize MyoXVIIIA. Furthermore, MyoXVIIIA but not MyoIIA localized to the surface of COS-1 cells, and most importantly, expression of MyoXVIIIA in COS-1 cells conferred SP-A binding. Western analysis of recombinant MyoXVIIIA domains expressed in bacteria mapped the epitopes of previously derived SP-R210 antibodies to the neck region of MyoXVIIIA. Antibodies raised against the neck domain of MyoXVIIIA blocked the binding of SP-A to macrophages. Together, these findings indicate that MyoXVIIIA constitutes a novel receptor for SP-A.

Received for publication, May 12, 2005 , and in revised form, July 11, 2005.

^* This work was supported by NHLBI Grants HL068127 (to Z. C. C.) and GM37537 (to D. F. H.) and NHLBI SCOR Grant HL56387 (to J. A. W.) from the National Institutes of Health and a Parker Francis Fellowship grant (to Z. C. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AAV80770, AY692137, AY692138, and AY692139.

¹ To whom correspondence should be addressed: University of Texas Health Center, Center of Biomedical Research, 11937 U. S. Highway 271, Tyler, TX 75708-3154. Tel.: 903-877-7941; Fax: 903-877-5876; E-mail: zissis.chroneos{at}uthct.edu.

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