TY - JOUR
T1 - Localization of glycosaminoglycan substitution sites on domain V of mouse perlecan.
AU - Tapanadechopone, P
AU - Hassell, J R
AU - Rigatti, B
AU - Couchman, J R
N1 - Keywords: Amino Acid Sequence; Animals; Base Sequence; Binding Sites; COS Cells; DNA Primers; Glycosaminoglycans; Glycosylation; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Mice; Molecular Sequence Data; Protein Structure, Tertiary; Proteoglycans; Rats; Recombinant Proteins; Sequence Homology, Amino Acid; Transfection
PY - 1999
Y1 - 1999
N2 - Perlecan, the predominant basement membrane proteoglycan, has previously been shown to contain glycosaminoglycans attached at serine residues, numbers 65, 71, and 76, in domain I. However, the C-terminal domains IV and V of this molecule may also be substituted with glycosaminoglycan chains, but the exact substitution sites were not identified. The amino acid sequence of mouse perlecan reveals many ser-gly sequences in these domains that are possible sites for glycosaminoglycan substitution. We expressed recombinant domain IV and/or V of mouse perlecan in COS-7 cells and analyzed glycosaminoglycan substitution. Both heparan sulfate and chondroitin sulfate chains could be detected on recombinant domain V. One site, ser-gly-glu (serine residue 3593), toward the C-terminal region of domain V is a substitution site for heparan sulfate. When this sequence was absent, chondroitin/dermatan sulfate substitution was deleted, and the likely site for this galactosaminoglycan substitution was ser-gly-ala-gly (serine residue 3250) on domain V.
AB - Perlecan, the predominant basement membrane proteoglycan, has previously been shown to contain glycosaminoglycans attached at serine residues, numbers 65, 71, and 76, in domain I. However, the C-terminal domains IV and V of this molecule may also be substituted with glycosaminoglycan chains, but the exact substitution sites were not identified. The amino acid sequence of mouse perlecan reveals many ser-gly sequences in these domains that are possible sites for glycosaminoglycan substitution. We expressed recombinant domain IV and/or V of mouse perlecan in COS-7 cells and analyzed glycosaminoglycan substitution. Both heparan sulfate and chondroitin sulfate chains could be detected on recombinant domain V. One site, ser-gly-glu (serine residue 3593), toward the C-terminal region of domain V is a substitution site for heparan sulfate. When this sequence was absent, chondroitin/dermatan sulfate substitution was deleted, and the likely site for this galactosaminoglycan substitution was ser-gly-ala-gly (serine residue 3250) on domain V.
U2 - 10.1006/bbrc.1999.1714
DO - 10.1006/bbrc.1999.1714
M3 - Journal article
C2 - 10600481
SN - 0006-291X
VL - 265
SP - 680
EP - 690
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -