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LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.

Banerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG - J. Cell Biol. (1999)

Bottom Line: Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA.However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels.Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

View Article: PubMed Central - PubMed

Affiliation: University of Oxford, Molecular Immunology Group, Nuffield Department of Medicine, John Radcliff Hospital, Headington, Oxford OX3 9DU, United Kingdom.

ABSTRACT
The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes where it is degraded before entering the circulation for rapid uptake by the liver. Currently, however, the identities of HA binding molecules which control this pathway are unknown. Here we describe the first such molecule, LYVE-1, which we have identified as a major receptor for HA on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single Link module the prototypic HA binding domain of the Link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

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Cell surface expression of LYVE-1 receptor on transfected COS cells. In A, COS 1 cells were transiently transfected with either full-length LYVE-1 cDNA in the expression vector pRcCMV (a and b), or with a control empty pRcCMV vector (c and d) using  DEAE dextran followed by surface immunofluorescent staining with rabbit polyclonal LYVE-1 antiserum (1:100 dilution) and FITC  goat anti–rabbit IgG. In B, control and LYVE-1 transfected COS cells were electrophoresed on a 10% polyacrylamide SDS-PAGE gel,  transferred to nitrocellulose, and Western blotted with LYVE-1 antiserum and peroxidase-conjugated goat anti–rabbit IgG (see Materials and Methods). Samples were control transfected COS (lane 1) and LYVE-1 transfected COS (lane 2). The positions and sizes in kilodaltons of the molecular mass calibration markers myosin (205 kD), β-galactosidase (116 kD), phosphorylase b (97 kD), BSA (66 kD),  ovalbumin (45 kD), and carbonic anhydrase (29 kD) are indicated on the left.
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Figure 3: Cell surface expression of LYVE-1 receptor on transfected COS cells. In A, COS 1 cells were transiently transfected with either full-length LYVE-1 cDNA in the expression vector pRcCMV (a and b), or with a control empty pRcCMV vector (c and d) using DEAE dextran followed by surface immunofluorescent staining with rabbit polyclonal LYVE-1 antiserum (1:100 dilution) and FITC goat anti–rabbit IgG. In B, control and LYVE-1 transfected COS cells were electrophoresed on a 10% polyacrylamide SDS-PAGE gel, transferred to nitrocellulose, and Western blotted with LYVE-1 antiserum and peroxidase-conjugated goat anti–rabbit IgG (see Materials and Methods). Samples were control transfected COS (lane 1) and LYVE-1 transfected COS (lane 2). The positions and sizes in kilodaltons of the molecular mass calibration markers myosin (205 kD), β-galactosidase (116 kD), phosphorylase b (97 kD), BSA (66 kD), ovalbumin (45 kD), and carbonic anhydrase (29 kD) are indicated on the left.

Mentions: To confirm the identification of the LYVE-1 molecule as an integral membrane glycoprotein, COS 1 fibroblasts were transfected with LYVE-1 cDNA and stained with polyclonal LYVE-1 antibodies for immunofluorescence microscopy. The specificity of the LYVE-1 serum is characterized in detail below. As shown by the results in Fig. 3 A, the transfectants displayed intense surface staining. Western blotting of the LYVE-1 transfected COS cells using a polyclonal serum generated against soluble LYVE-1 revealed a major band which migrated with an apparent molecular mass of 60 kD, and which was absent from mock transfected cells. The discrepancy between the experimentally observed value and the size predicted from the primary sequence (35,158 D) is largely due to glycosylation, since treatment with either N-glycanase or O-glycanase/neuraminidase reduced the molecular mass by ∼20 kD (not shown).


LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan.

Banerji S, Ni J, Wang SX, Clasper S, Su J, Tammi R, Jones M, Jackson DG - J. Cell Biol. (1999)

Cell surface expression of LYVE-1 receptor on transfected COS cells. In A, COS 1 cells were transiently transfected with either full-length LYVE-1 cDNA in the expression vector pRcCMV (a and b), or with a control empty pRcCMV vector (c and d) using  DEAE dextran followed by surface immunofluorescent staining with rabbit polyclonal LYVE-1 antiserum (1:100 dilution) and FITC  goat anti–rabbit IgG. In B, control and LYVE-1 transfected COS cells were electrophoresed on a 10% polyacrylamide SDS-PAGE gel,  transferred to nitrocellulose, and Western blotted with LYVE-1 antiserum and peroxidase-conjugated goat anti–rabbit IgG (see Materials and Methods). Samples were control transfected COS (lane 1) and LYVE-1 transfected COS (lane 2). The positions and sizes in kilodaltons of the molecular mass calibration markers myosin (205 kD), β-galactosidase (116 kD), phosphorylase b (97 kD), BSA (66 kD),  ovalbumin (45 kD), and carbonic anhydrase (29 kD) are indicated on the left.
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Related In: Results  -  Collection

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Figure 3: Cell surface expression of LYVE-1 receptor on transfected COS cells. In A, COS 1 cells were transiently transfected with either full-length LYVE-1 cDNA in the expression vector pRcCMV (a and b), or with a control empty pRcCMV vector (c and d) using DEAE dextran followed by surface immunofluorescent staining with rabbit polyclonal LYVE-1 antiserum (1:100 dilution) and FITC goat anti–rabbit IgG. In B, control and LYVE-1 transfected COS cells were electrophoresed on a 10% polyacrylamide SDS-PAGE gel, transferred to nitrocellulose, and Western blotted with LYVE-1 antiserum and peroxidase-conjugated goat anti–rabbit IgG (see Materials and Methods). Samples were control transfected COS (lane 1) and LYVE-1 transfected COS (lane 2). The positions and sizes in kilodaltons of the molecular mass calibration markers myosin (205 kD), β-galactosidase (116 kD), phosphorylase b (97 kD), BSA (66 kD), ovalbumin (45 kD), and carbonic anhydrase (29 kD) are indicated on the left.
Mentions: To confirm the identification of the LYVE-1 molecule as an integral membrane glycoprotein, COS 1 fibroblasts were transfected with LYVE-1 cDNA and stained with polyclonal LYVE-1 antibodies for immunofluorescence microscopy. The specificity of the LYVE-1 serum is characterized in detail below. As shown by the results in Fig. 3 A, the transfectants displayed intense surface staining. Western blotting of the LYVE-1 transfected COS cells using a polyclonal serum generated against soluble LYVE-1 revealed a major band which migrated with an apparent molecular mass of 60 kD, and which was absent from mock transfected cells. The discrepancy between the experimentally observed value and the size predicted from the primary sequence (35,158 D) is largely due to glycosylation, since treatment with either N-glycanase or O-glycanase/neuraminidase reduced the molecular mass by ∼20 kD (not shown).

Bottom Line: Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA.However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels.Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

View Article: PubMed Central - PubMed

Affiliation: University of Oxford, Molecular Immunology Group, Nuffield Department of Medicine, John Radcliff Hospital, Headington, Oxford OX3 9DU, United Kingdom.

ABSTRACT
The extracellular matrix glycosaminoglycan hyaluronan (HA) is an abundant component of skin and mesenchymal tissues where it facilitates cell migration during wound healing, inflammation, and embryonic morphogenesis. Both during normal tissue homeostasis and particularly after tissue injury, HA is mobilized from these sites through lymphatic vessels to the lymph nodes where it is degraded before entering the circulation for rapid uptake by the liver. Currently, however, the identities of HA binding molecules which control this pathway are unknown. Here we describe the first such molecule, LYVE-1, which we have identified as a major receptor for HA on the lymph vessel wall. The deduced amino acid sequence of LYVE-1 predicts a 322-residue type I integral membrane polypeptide 41% similar to the CD44 HA receptor with a 212-residue extracellular domain containing a single Link module the prototypic HA binding domain of the Link protein superfamily. Like CD44, the LYVE-1 molecule binds both soluble and immobilized HA. However, unlike CD44, the LYVE-1 molecule colocalizes with HA on the luminal face of the lymph vessel wall and is completely absent from blood vessels. Hence, LYVE-1 is the first lymph-specific HA receptor to be characterized and is a uniquely powerful marker for lymph vessels themselves.

Show MeSH
Related in: MedlinePlus