PMID- 10581249
OWN - NLM
STAT- MEDLINE
DCOM- 20000131
LR  - 20190406
IS  - 0261-4189 (Print)
IS  - 0261-4189 (Linking)
VI  - 18
IP  - 23
DP  - 1999 Dec 1
TI  - Interaction of agrin with laminin requires a coiled-coil conformation of the
      agrin-binding site within the laminin gamma1 chain.
PG  - 6762-70
AB  - Coiled-coil domains are found in a wide variety of proteins, where they typically
      specify subunit oligomerization. Recently, we have demonstrated that agrin, a
      multidomain heparan sulfate proteoglycan with a crucial role in the development
      of the nerve-muscle synapse, binds to the three-stranded coiled-coil domain of
      laminin-1. The interaction with laminin mediates the integration of agrin into
      basement membranes. Here we characterize the binding site within the laminin-1
      coiled coil in detail. Binding assays with individual laminin-1 full-length
      chains and fragments revealed that agrin specifically interacts with the gamma1
      subunit of laminin-1, whereas no binding to alpha1 and beta1 chains was detected.
      By using recombinant gamma1 chain fragments, we mapped the binding site to a
      sequence of 20 residues. Furthermore, we demonstrate that a coiled-coil
      conformation of this binding site is required for its interaction with agrin. The
      finding that recombinant gamma1 fragments bound at least 10-fold less than native
      laminin-1 indicates that the structure of the three-stranded coiled-coil domain
      of laminin is required for high-affinity agrin binding. Interestingly, no binding
      to a chimeric gamma2 fragment was observed, indicating that the interaction of
      agrin with laminin is isoform specific.
FAU - Kammerer, R A
AU  - Kammerer RA
AD  - Departments of Biophysical Chemistry, Biozentrum, University of Basel,
      Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
FAU - Schulthess, T
AU  - Schulthess T
FAU - Landwehr, R
AU  - Landwehr R
FAU - Schumacher, B
AU  - Schumacher B
FAU - Lustig, A
AU  - Lustig A
FAU - Yurchenco, P D
AU  - Yurchenco PD
FAU - Ruegg, M A
AU  - Ruegg MA
FAU - Engel, J
AU  - Engel J
FAU - Denzer, A J
AU  - Denzer AJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - EMBO J
JT  - The EMBO journal
JID - 8208664
RN  - 0 (Agrin)
RN  - 0 (DNA, Complementary)
RN  - 0 (Laminin)
RN  - 0 (Recombinant Proteins)
RN  - 0 (laminin 1)
SB  - IM
MH  - Agrin/*chemistry/*metabolism
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - COS Cells
MH  - Circular Dichroism
MH  - DNA, Complementary/metabolism
MH  - Escherichia coli/metabolism
MH  - Gene Deletion
MH  - Laminin/*chemistry/genetics/*metabolism
MH  - Molecular Sequence Data
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Temperature
MH  - Transfection
MH  - Ultracentrifugation
PMC - PMC1171738
EDAT- 1999/12/03 00:00
MHDA- 1999/12/03 00:01
CRDT- 1999/12/03 00:00
PHST- 1999/12/03 00:00 [pubmed]
PHST- 1999/12/03 00:01 [medline]
PHST- 1999/12/03 00:00 [entrez]
AID - 10.1093/emboj/18.23.6762 [doi]
PST - ppublish
SO  - EMBO J. 1999 Dec 1;18(23):6762-70. doi: 10.1093/emboj/18.23.6762.