Partial amino acid sequence of human thrombospondin as determined by analysis of cDNA clones: homology to malarial circumsporozoite proteins

• Published in: Biochemistry (Easton) 1986-12, Vol.25 (26), p.8418-8425
• Main Authors: Kobayashi, Sentaro, Eden-McCutchan, Francine, Framson, Paul, Bornstein, Paul
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antigens, Protozoan - genetics; Antigens, Surface - genetics; Base Sequence; Biological and medical sciences; Blood coagulation. Blood cells; Cloning, Molecular; DNA - analysis; DNA Restriction Enzymes; endothelium; Endothelium - metabolism; Fundamental and applied biological sciences. Psychology; Glycoproteins - genetics; Humans; man; Molecular and cellular biology; Nucleic Acid Hybridization; nucleotide sequence; Plasmodium - genetics; Plasmodium falciparum - genetics; Platelet; Protozoan Proteins; Sequence Homology, Nucleic Acid; thrombospondin; Thrombospondins; Umbilical Veins - metabolism
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00374a014

A lambda gt 11 library prepared from human umbilical vein endothelial cell RNA was screened for cDNAs encoding thrombospondin. Reagents included a monospecific antibody to human thrombospondin and a mixture of four synthetic oligodeoxyribonucleotides derived from an amino acid sequence near the NH2 terminus of mature human thrombospondin. Two series of cDNA clones coding for sequences at the 5' and 3' ends of thrombospondin mRNA, respectively, were isolated. The nucleotide sequence of a 1.3-kilobase (kb) 5' clone (lambda TS-33) coded for 99 bases of 5' untranslated RNA, a signal peptide of 18 amino acids, and the first 379 amino acids of thrombospondin. Northern blot analysis with lambda TS-33 detected a single mRNA species of approximately 6.0 kb in rat aortic smooth muscle cell RNA. Thrombospondin mRNA levels increased rapidly, but transiently, in quiescent smooth muscle cells treated with platelet-derived growth factor. The kinetics of this response were very similar to those of the thrombospondin protein to this growth factor. There was significant homology in amino acid sequence between thrombospondin and a conserved region in the circumsporozoite protein of two malarial sporozoites. This region of thrombospondin may therefore represent a potential recognition site for a cell surface thrombospondin receptor.