title: EMILIN, a component of the elastic fiber and a new member of the C1q/tumor necrosis factor superfamily of proteins.
abstract: EMILIN (elastin microfibril interface located protein) is an extracellular matrix glycoprotein abundantly expressed in elastin-rich tissues such as blood vessels, skin, heart, and lung. It occurs associated with elastic fibers at the interface between amorphous elastin and microfibrils. Avian EMILIN was extracted from 19-day-old embryonic chick aortas and associated blood vessels and purified by ion-exchange chromatography and gel filtration. Tryptic peptides were generated from EMILIN and sequenced, and degenerate inosine-containing oligonucleotide primers were designed from some peptides. A set of primers allowed the amplification of a 360-base pair reverse transcription polymerase chain reaction product from chick aorta mRNA. A probe based on a human homologue selected by comparison of the chick sequence with EST data base was used to select overlapping clones from both human aorta and kidney cDNA libraries. Here we present the cDNA sequence of the entire coding region of human EMILIN encompassing an open reading frame of 1016 amino acid residues. There was a high degree of homology (76% identity and 88% similarity) between the chick C terminus and the human sequence as well as between the N terminus of the mature chick protein where 10 of 12 residues, as determined by N-terminal sequencing, were identical or similar to the deduced N terminus of human EMILIN. The domain organization of human EMILIN includes a C1q-like globular domain at the C terminus, a collagenous stalk, and a longer segment in which at least four heptad repeats and a leucine zipper can be identified with a high potential for forming coiled-coil alpha helices. At the N terminus there is a cysteine-rich sequence stretch similar to a region of multimerin, a platelet and endothelial cell component, containing a partial epidermal growth factor-like motif. The native state of the recombinantly expressed EMILIN C1q-like domain to be used in cell adhesion was determined by CD spectra analysis, which indicated a high value of beta-sheet conformation. The EMILIN C1q-like domain promoted a high cell adhesion of the leiomyosarcoma cell line SK-UT-1, whereas the fibrosarcoma cell line HT1080 was negative.
authors.affiliation: Divisione di Oncologia Sperimentale 2, Centro di Riferimento Oncologico di Aviano, 33081 Aviano, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Doliana
authors.forename: M
authors.initials: M
authors.lastname: Mongiat
authors.forename: F
authors.initials: F
authors.lastname: Bucciotti
authors.forename: E
authors.initials: E
authors.lastname: Giacomello
authors.forename: R
authors.initials: R
authors.lastname: Deutzmann
authors.forename: D
authors.initials: D
authors.lastname: Volpin
authors.forename: G M
authors.initials: GM
authors.lastname: Bressan
authors.forename: A
authors.initials: A
authors.lastname: Colombatti
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000737:chemistry; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D002642:Chick Embryo; D002942:Circular Dichroism; D015922:Complement C1q; D018076:DNA, Complementary / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D015723:Gene Library; D006801:Humans; D016350:Leucine Zippers; D008562:Membrane Glycoproteins / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000145:classification / Q000302:isolation & purification; D020449:Repetitive Sequences, Amino Acid; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014409:Tumor Necrosis Factor-alpha
chemical_list: D015815:Cell Adhesion Molecules; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C030340:elastin microfibril interface located protein; D015922:Complement C1q