Identification of cDNA clones encoding different domains of the basement membrane heparan sulfate proteoglycan.
We have used antibodies to the basement membrane proteoglycan to screen lambda gt11 expression vector libraries and have isolated two cDNA clones, termed BPG 5 and BPG 7, which encode different portions of the core protein of the heparan sulfate basement membrane proteoglycan. These clones hybridize to a single mRNA species of approximately 12 kilobases. Amino acid sequences obtained on peptides derived from protease digests of the core protein were found in the deduced sequence, confirming the identity of these clones. BPG 5 spanned 1986 base pairs and has an open reading frame of 662 amino acids. The amino acid sequence deduced from BPG 5 contains two cysteine-rich domains and two internally homologous domains lacking cysteine. The cysteine-rich domains show homology to the cysteine-rich domains of the laminin chains. A globule-rod structure, similar to that of the short arms of the laminin chains, is proposed for this region of the proteoglycan. The other clone, BPG 7, is 2193 base pairs long and has an open reading frame of 731 amino acids. The deduced sequence contains eight internal repeats with 2 cysteine residues in each repeat. These repeats show homology to the neural-cell adhesion molecule N-CAM and the plasma alpha 1B-glycoprotein. Looping structures similar to these proteins and to other proteins of the immunoglobulin gene superfamily are proposed for this region of the proteoglycan. The sequence DSGEY was found four times in this domain and could be heparan sulfate attachment sites.
Noonan
D M
DM
Laboratory of Developmental Biology and Anomalies, National Institute of Dental Research, Bethesda, Maryland 20892.
Horigan
E A
EA
Ledbetter
S R
SR
Vogeli
G
G
Sasaki
M
M
Yamada
Y
Y
Hassell
J R
JR
D000595:Amino Acid Sequence; D001485:Basement Membrane / Q000378:metabolism*; D015152:Blotting, Northern; D002460:Cell Line; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005796:Genes; D005822:Genetic Vectors; D006025:Glycosaminoglycans / Q000235:genetics*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000235:genetics*; D007797:Laminin / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D011487:Protein Conformation; D011509:Proteoglycans / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D011508:Chondroitin Sulfate Proteoglycans; D004247:DNA; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D007797:Laminin; D010446:Peptide Fragments; D011509:Proteoglycans