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

VL - 265

SP - 680

EP - 690

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 3

ER -