Identification and characterization of CPAMD8, a novel member of the complement 3/α2-macroglobulin family with a C-terminal Kazal domain☆
Section snippets
Identification and cloning of CPAMD8
Using the six known protein sequences of the C3/α2M family members, we performed a homologue search of the human genome database with the tBLASTn program. Seven additional putative C3/α2M-like genes were found in three genomic contigs on chromosomes 7, 12, and 19 (Table 1). Four putative genes on chromosome 12 are clustered with α2M and PZP. CPAMD8 is located on chromosome 19 (19p13.2–p13.3). A putative cDNA was predicted with GENESCAN, consists of 40 exons (∼6 kb), and has significant
Discussion
We report the cloning and characterization of a novel member of the C3/α2M family. CPAMD8 is highly conserved during evolution from zebrafish to human. CPAMD8 orthologues showing 65–86% amino acid identity are present in several species such as pig, cow, chicken, Fugu, zebrafish, and Xenopus. Interestingly, there is no evidence that CPAMD8 exists in rodents. The PAR4 gene, which overlaps with CPAMD8 in the human genome, has been found on mouse chromosome 8, but CPAMD8 is absent from that
Materials
Superscript one-step RT-PCR, Lipofectamine 2000, pSecTag2B vector, and Precast gels for SDS–PAGE were purchased from Invitrogen (Carlsbad, CA, USA). The PROTEIN Script II Linked Transcription:Translation kit was purchased form Ambion (Austin, TX, USA). The Redivue l-[35S]methionine, pGEX-4T-1 vector, glutathione Sepharose 4B, and ECL Western blotting detect reagent were purchased from Amersham Pharmacia Biotech (Piscataway, NJ, USA). Centricon centrifugal filter devices were from Millipore
Acknowledgements
We thank the Kazusa DNA Research Institute for the partial cDNA clone of CPAMD8, Tristan Williams for help with mass spectrometry, Helene Fisher and Darek Kedra for advice, and Torgny Stigbrand for comments on the manuscript. This research was supported by a grant from the National Institutes of Health to E.E.
References (50)
α2-Macroglobulin and related thiol ester plasma proteins
α2-Macroglobulin: an evolutionarily conserved arm of the innate immune system
Dev. Comp. Immunol
(1999)Cell surface antigen CD109 is a novel member of the alpha (2) macroglobulin/C3, C4, C5 family of thioester-containing proteins
Blood
(2002)- et al.
C3 component of complement secreted by established cell lines
Cell
(1978) - et al.
Human pregnancy zone protein and alpha 2-macroglobulin: high-affinity binding of complexes to the same receptor on fibroblasts and characterization by monoclonal antibodies
J. Biol. Chem
(1986) - et al.
Domain structure of human alpha 2-macroglobulin: characterization of a receptor-binding domain obtained by digestion with papain
FEBS Lett
(1986) - et al.
Classification of alpha 2-macroglobulin–cytokine interactions based on affinity of noncovalent association in solution under apparent equilibrium conditions
J. Biol. Chem
(1994) Identification of the high affinity binding site in transforming growth factor-beta involved in complex formation with alpha 2-macroglobulin: implications regarding the molecular mechanisms of complex formation between alpha 2-macroglobulin and growth factors, cytokines, and hormones
J. Biol. Chem
(2001)Identical or overlapping sequences in the primary structure of human alpha(2)-macroglobulin are responsible for the binding of nerve growth factor-beta, platelet-derived growth factor-BB, and transforming growth factor-beta
J. Biol. Chem
(2000)- et al.
Aberrant forms of alpha(2)-macroglobulin purified from patients with multiple sclerosis
Clin. Chim. Acta
(2000)
Identification of a cell-surface antigen associated with activated T lymphoblasts and activated platelets
Blood
CD109 is expressed on a subpopulation of CD34+ cells enriched in hematopoietic stem and progenitor cells
Exp. Hematol
Expansion of genes encoding complement components in bony fish: biological implications of the complement diversity
Dev. Comp. Immunol
Murinoglobulin, a novel protease inhibitor from murine plasma: isolation, characterization, and comparison with murine alpha-macroglobulin and human alpha-2-macroglobulin
J. Biol. Chem
Identification of four genes coding for isoforms of murinoglobulin, the monomeric mouse alpha 2-macroglobulin: characterization of the exons coding for the bait region
Genomics
Conserved role of a complement-like protein in phagocytosis revealed by dsDNA knockout in cultured cells of the mosquito, Anopheles gambiae
Cell
Protein prenylation: a pivotal posttranslational process
Biochem. Biophys. Res. Commun
NF-κB mediated IL-1β-induced synthesis/release of α2-macroglobulin in a human glial cell line
Mol. Brain Res
The structure of the bovine pancreatic secretory trypsin inhibitor–Kazal's inhibitor: II. The order of the tryptic peptides
J. Biol. Chem
Crystal structures of the heparan sulphate-binding domain of follistatin: insights into ligand binding
J. Biol. Chem
Prediction of complete gene structures in human genomic DNA
J. Mol. Biol
Identification of a novel family of cell-surface proteins expressed in human vascular endothelium
J. Biol. Chem
Determination of integrins on cells by cell adhesion to antibodies
Anal. Biochem
The role of complement in innate and adaptive immunity
Curr. Top. Microbiol. Immunol
Biosynthesis and genetics of C3
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Sequence data from this article have been deposited with the NCBI Data Library under the Accession No. AY101765. The amino acid sequence of this protein can be accessed through the NCBI protein database under NCBI Accession No. AAM50084.