Figure 1. Protein Lunapark is an N-myristoylated integral membrane protein.

A. Structure and N-terminal sequence of cDNA clones analyzed in this study. B. mRNAs encoding ARF1-, SERINC1-, KIAA1609-, Lunapark- and ZZEF1-FLAG were translated in vitro in the presence of [³H]leucine ([³H]Leu) or [³H]myristic acid ([³H]Myr) using an insect cell-free protein synthesis system. The labeled translation products were analyzed by SDS-PAGE and fluorography. C. cDNAs encoding ARF1-, SERINC1-, KIAA1609-, Lunapark- and ZZEF1-FLAG were transfected in to COS-1 cells, and their expression and N-myristoylation of the products were evaluated by Western blotting analysis and [³H]myristic acid labeling, respectively. D. KIAA1609-FLAG and Lunapark-FLAG were expressed in COS-1 cells, and subcellular fractionation experiments were performed. Hsp70 was used as a cytosolic marker protein and PDI was used as membrane/organelle marker protein. C, cytosolic fraction; M, membrane/organelle fraction; N, nuclear fraction.