Cross-linking of laminin-nidogen complexes by tissue transglutaminase. A novel mechanism for basement membrane stabilization.
The laminin-nidogen complex, a major component of basement membranes, incorporates [3H]putrescine and monodansylcadaverine in the presence of guinea pig liver transglutaminase. Label was detected in nidogen in the isolated, as well as in the complexed form, but not in laminin. The incorporation proceeds in a time-dependent manner at a rate similar to that achieved with N,N-dimethylcasein, a well characterized transglutaminase substrate. Saturation of incorporation site(s), as well as comparison with the incorporation level in reference proteins, indicated the presence of one high affinity amine acceptor site in nidogen. Electron microscopy of the reaction products showed that the laminin-nidogen complexes become stabilized in a head-to-head arrangement, characteristic of Ca(2+)-induced self-aggregation. Indirect immunofluorescence and detection of transglutaminase activity on unfixed cryosections revealed an extracellular distribution of tissue transglutaminase. Intensive staining was observed in collagen-rich connective tissue. Codistribution with nidogen was not a ubiquitous feature, but was observed in many locations.
M. E. Müller-Institute for Biomechanics, University of Bern, Switzerland.
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Aeschlimann
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Paulsson
D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D015153:Blotting, Western; D002103:Cadaverine / Q000031:analogs & derivatives / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006168:Guinea Pigs; D007668:Kidney / Q000378:metabolism; D007797:Laminin / Q000378:metabolism* / Q000648:ultrastructure; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D009206:Myocardium / Q000378:metabolism; D011700:Putrescine / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism*
D005353:Fibronectins; D005456:Fluorescent Dyes; D007797:Laminin; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C029967:nidogen; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine; D011700:Putrescine