PMID- 10364234
OWN - NLM
STAT- MEDLINE
DCOM- 19990715
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 25
DP  - 1999 Jun 18
TI  - Mammalian homologues of the Drosophila slit protein are ligands of the heparan
      sulfate proteoglycan glypican-1 in brain.
PG  - 17885-92
AB  - Using an affinity matrix in which a recombinant glypican-Fc fusion protein
      expressed in 293 cells was coupled to protein A-Sepharose, we have isolated from 
      rat brain at least two proteins that were detected by SDS-polyacrylamide gel
      electrophoresis as a single 200-kDa silver-stained band, from which 16 partial
      peptide sequences were obtained by nano-electrospray tandem mass spectrometry.
      Mouse expressed sequence tags containing two of these peptides were employed for 
      oligonucleotide design and synthesis of probes by polymerase chain reaction and
      enabled us to isolate from a rat brain cDNA library a 4.1-kilobase clone that
      encoded two of our peptide sequences and represented the N-terminal portion of a 
      protein containing a signal peptide and three leucine-rich repeats. Comparisons
      with recently published sequences also showed that our peptides were derived from
      proteins that are members of the Slit/MEGF protein family, which share a number
      of structural features such as N-terminal leucine-rich repeats and C-terminal
      epidermal growth factor-like motifs, and in Drosophila Slit is necessary for the 
      development of midline glia and commissural axon pathways. All of the five known 
      rat and human Slit proteins contain 1523-1534 amino acids, and our peptide
      sequences correspond best to those present in human Slit-1 and Slit-2. Binding of
      these ligands to the glypican-Fc fusion protein requires the presence of the
      heparan sulfate chains, but the interaction appears to be relatively specific for
      glypican-1 insofar as no other identified heparin-binding proteins were isolated 
      using our affinity matrix. Northern analysis demonstrated the presence of two
      mRNA species of 8. 6 and 7.5 kilobase pairs using probes based on both N- and
      C-terminal sequences, and in situ hybridization histochemistry showed that these 
      glypican-1 ligands are synthesized by neurons, such as hippocampal pyramidal
      cells and cerebellar granule cells, where we have previously also demonstrated
      glypican-1 mRNA and immunoreactivity. Our results therefore indicate that Slit
      family proteins are functional ligands of glypican-1 in nervous tissue and
      suggest that their interactions may be critical for certain stages of central
      nervous system histogenesis.
FAU - Liang, Y
AU  - Liang Y
AD  - Department of Pharmacology, New York University School of Medicine, New York, New
      York 10016, USA.
FAU - Annan, R S
AU  - Annan RS
FAU - Carr, S A
AU  - Carr SA
FAU - Popp, S
AU  - Popp S
FAU - Mevissen, M
AU  - Mevissen M
FAU - Margolis, R K
AU  - Margolis RK
FAU - Margolis, R U
AU  - Margolis RU
LA  - eng
SI  - GENBANK/AF141386
GR  - MH-00129/MH/NIMH NIH HHS/United States
GR  - NS-09348/NS/NINDS NIH HHS/United States
GR  - NS-13876/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Drosophila Proteins)
RN  - 0 (Heparan Sulfate Proteoglycans)
RN  - 0 (Ligands)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (sli protein, Drosophila)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Brain/metabolism
MH  - Cloning, Molecular
MH  - Drosophila/*metabolism
MH  - *Drosophila Proteins
MH  - Heparan Sulfate Proteoglycans/*metabolism
MH  - Humans
MH  - *Ligands
MH  - Mice
MH  - Molecular Sequence Data
MH  - Nerve Tissue Proteins/*chemistry/genetics
MH  - Peptide Fragments/chemistry
MH  - RNA, Messenger
MH  - Rats
MH  - Recombinant Fusion Proteins/genetics
MH  - Sequence Alignment
EDAT- 1999/06/11 00:00
MHDA- 1999/06/11 00:01
CRDT- 1999/06/11 00:00
PHST- 1999/06/11 00:00 [pubmed]
PHST- 1999/06/11 00:01 [medline]
PHST- 1999/06/11 00:00 [entrez]
AID - 10.1074/jbc.274.25.17885 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Jun 18;274(25):17885-92. doi: 10.1074/jbc.274.25.17885.