PMID- 10593924
OWN - NLM
STAT- MEDLINE
DCOM- 20000127
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 51
DP  - 1999 Dec 17
TI  - The integrin alpha(9)beta(1) binds to a novel recognition sequence (SVVYGLR) in
      the thrombin-cleaved amino-terminal fragment of osteopontin.
PG  - 36328-34
AB  - The integrin alpha(9)beta(1) mediates cell adhesion to tenascin-C and VCAM-1 by
      binding to sequences distinct from the common integrin-recognition sequence,
      arginine-glycine-aspartic acid (RGD). A thrombin-cleaved NH(2)-terminal fragment 
      of osteopontin containing the RGD sequence has recently been shown to also be a
      ligand for alpha(9)beta(1). In this report, we used site-directed mutagenesis and
      synthetic peptides to identify the alpha(9)beta(1) recognition sequence in
      osteopontin. alpha(9)-transfected SW480, Chinese hamster ovary, and L-cells
      adhered to a recombinant NH(2)-terminal osteopontin fragment in which the RGD
      site was mutated to RAA (nOPN-RAA). Adhesion was completely inhibited by
      anti-alpha(9) monoclonal antibody Y9A2, indicating the presence of a non-RGD
      alpha(9)beta(1) recognition sequence within this fragment. Alanine substitution
      mutagenesis of 13 additional conserved negatively charged amino acid residues in 
      this fragment had no effect on alpha(9)beta(1)-mediated adhesion, but adhesion
      was dramatically inhibited by either alanine substitution or deletion of tyrosine
      165. A synthetic peptide, SVVYGLR, corresponding to the sequence surrounding
      Tyr(165), blocked alpha(9)beta(1)-mediated adhesion to nOPN-RAA and exposed a
      ligand-binding-dependent epitope on the integrin beta(1) subunit on
      alpha(9)-transfected, but not on mock-transfected cells. These results
      demonstrate that the linear sequence SVVYGLR directly binds to alpha(9)beta(1)
      and is responsible for alpha(9)beta(1)-mediated cell adhesion to the
      NH(2)-terminal fragment of osteopontin.
FAU - Yokosaki, Y
AU  - Yokosaki Y
AD  - Department of Internal Medicine, National Hiroshima Hospital, 513 Jike, Saijoh,
      Higashi-Hiroshima 739-0041, Japan. yokosaki@hiroshima.hosp.go.jp
FAU - Matsuura, N
AU  - Matsuura N
FAU - Sasaki, T
AU  - Sasaki T
FAU - Murakami, I
AU  - Murakami I
FAU - Schneider, H
AU  - Schneider H
FAU - Higashiyama, S
AU  - Higashiyama S
FAU - Saitoh, Y
AU  - Saitoh Y
FAU - Yamakido, M
AU  - Yamakido M
FAU - Taooka, Y
AU  - Taooka Y
FAU - Sheppard, D
AU  - Sheppard D
LA  - eng
GR  - HL/AI33259/HL/NHLBI NIH HHS/United States
GR  - HL47412/HL/NHLBI NIH HHS/United States
GR  - HL53949/HL/NHLBI NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
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 (Integrins)
RN  - 0 (SPP1 protein, human)
RN  - 0 (Sialoglycoproteins)
RN  - 0 (integrin alpha 9 beta 1)
RN  - 106441-73-0 (Osteopontin)
RN  - EC 3.4.21.5 (Thrombin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Binding Sites/genetics
MH  - CHO Cells
MH  - Cricetinae
MH  - Humans
MH  - Integrins/chemistry/genetics/*metabolism
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Osteopontin
MH  - Protein Binding
MH  - Sialoglycoproteins/chemistry/genetics/*metabolism
MH  - Thrombin/metabolism
MH  - Transfection
MH  - Tumor Cells, Cultured
EDAT- 1999/12/14 00:00
MHDA- 1999/12/14 00:01
CRDT- 1999/12/14 00:00
PHST- 1999/12/14 00:00 [pubmed]
PHST- 1999/12/14 00:01 [medline]
PHST- 1999/12/14 00:00 [entrez]
AID - 10.1074/jbc.274.51.36328 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Dec 17;274(51):36328-34. doi: 10.1074/jbc.274.51.36328.