Isolation, characterization and localization of a lectin within the vitelline membrane of the hen's egg.
A lectin with an affinity for certain sulphated polysaccharides, such as fucoidin and dextran sulphate, has been isolated from the vitelline membrane of hens' eggs and purified to homogeneity as assessed by two-dimensional gel electrophoresis. Polyclonal and monoclonal antibodies have been raised to the lectin and used in indirect immunofluorescence microscopy to localize the agglutinin in the outer layer of the vitelline membrane, where the lectin persists prior to the breakdown of the vitelline membrane. The quantity of lectin extracted from the two layers of the membrane, which have been separated by the method of Bellairs, Harkness & Harkness (1963), correlated well with the results of immunofluorescence microscopy. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of the two layers of the membrane indicates that each layer has a distinctive polypeptide composition, the outer layer containing in particular lysozyme and avidin. The evidence obtained in this study indicates that the lectin is not involved in adhesion of the blastoderm to the vitelline membrane; neither is it involved in the expression of the blastoderm nor in maintaining the strength of the membrane. The possible roles in promoting transport of solutes across the membrane as well as providing bactericidal properties to the egg are discussed.
G M
GM
Cook
R
R
Bellairs
N G
NG
Rutherford
C A
CA
Stafford
T
T
Alderson
D000818:Animals; D002645:Chickens; D016264:Dextran Sulfate; D003911:Dextrans / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006384:Hemagglutination / Q000187:drug effects; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D037102:Lectins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010063:Ovum / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D011134:Polysaccharides / Q000494:pharmacology; D014817:Vitelline Membrane / Q000032:analysis* / Q000276:immunology
D003911:Dextrans; D037102:Lectins; D010455:Peptides; D011134:Polysaccharides; D016264:Dextran Sulfate; C007789:fucoidan