Galectin-3 is associated with the plasma membrane of lens fiber cells.
OBJECTIVE
To discover proteins that have the potential to contribute to the tight packing of fiber cells in the lens.


METHODS
Crude fiber cell membranes were isolated from ovine lens cortex. Proteins were separated by two-dimensional gel electrophoresis, and selected protein spots identified by micro-sequencing. The identification of galectin-3 was confirmed by immunoblotting with a specific antibody. The association of galectin-3 with the fiber cell plasma membrane was investigated using immunofluorescence microscopy, solubilization trials with selected reagents, and immunoprecipitation to identify candidate ligands.


RESULTS
A cluster of three protein spots with an apparent molecular weight of 31,000 and isoelectric points ranging between 7 and 8.5 were resolved and identified as galectin-3. This protein was associated peripherally with the fiber cell plasma membrane and interacted with MP20, an abundant intrinsic membrane protein that had been identified previously as a component of membrane junctions between fiber cells.


CONCLUSIONS
The detection of galectin-3 in the lens is a novel result and adds to the growing list of lens proteins with adhesive properties. Its location at the fiber cell membrane and its association with the junction-forming MP20 is consistent with a potential role in the development or maintenance of the tightly packed lens tissue architecture.
Gonen
T
T
School of Biological Sciences, University of Auckland, New Zealand.
Donaldson
P
P
Kistler
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000032:analysis*; D002462:Cell Membrane / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005136:Eye Proteins / Q000032:analysis*; D037502:Galectin 3; D015151:Immunoblotting; D007904:Lens Cortex, Crystalline / Q000737:chemistry* / Q000166:cytology; D008024:Ligands; D008562:Membrane Glycoproteins / Q000032:analysis*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D012756:Sheep
D000943:Antigens, Differentiation; D005136:Eye Proteins; D037502:Galectin 3; D008024:Ligands; D008562:Membrane Glycoproteins