Identification and characterization of tropomodulin and tropomyosin in the adult rat lens.
The lens fiber cells express all the major components of the erythrocyte membrane skeleton including spectrin, protein 4.1 and ankyrin. We have used immunoblot and immunoprecipitation analyses, as well as immunofluorescence localization to identify and characterize two additional components of the membrane skeleton in the rat lens: tropomyosin and the tropomyosin-binding protein tropomodulin. In the erythrocyte, tropomyosin and tropomodulin are proposed to stabilize and limit the lengths of the short actin filaments of the spectrin-actin network, thus influencing the organization and mechanical properties of the erythrocyte membrane skeleton. Antibodies directed against erythrocyte tropomodulin specifically recognize a M(r) 43,000 polypeptide from rat lens that comigrates with erythrocyte tropomodulin on SDS-gels. A non-muscle isoform of tropomyosin is also present in the lens. This tropomyosin isoform migrates on SDS-gels with a M(r) of approximately 28,000 and is distinct from the two erythrocyte isoforms of tropomyosin (M(r) 27,000 and 29,000). Indirect immunofluorescence staining of 5 microns cryosections of adult rat lens reveals that both tropomodulin and tropomyosin colocalize with rhodamine phalloidin staining for actin filaments on fiber cell plasma membranes. Lens tropomodulin exhibits many characteristics that are similar to its erythrocyte counterpart. For example, lens tropomodulin binds tropomyosin in a solid-phase blot binding assay, and extraction experiments with Triton X-100, urea and NaOH show that the membrane-bound tropomodulin in the lens is a tightly associated peripheral membrane protein that is a component of the Triton-insoluble cytoskeleton. However, unlike the erythrocyte, there are approximately 2000 actin monomers per tropomodulin in the lens.(ABSTRACT TRUNCATED AT 250 WORDS)
Woo
M K
MK
Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037.
Fowler
V M
VM
D000199:Actins / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007908:Lens, Crystalline / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D008840:Microfilament Proteins; D008970:Molecular Weight; D051381:Rats; D011917:Rats, Inbred Lew; D012995:Solubility; D013347:Subcellular Fractions / Q000378:metabolism; D051348:Tropomodulin; D014335:Tropomyosin / Q000302:isolation & purification / Q000378:metabolism*
D000199:Actins; D002352:Carrier Proteins; D008840:Microfilament Proteins; C495832:Tmod1 protein, rat; D051348:Tropomodulin; D014335:Tropomyosin