Tropomyosin isoform 5b is expressed in human erythrocytes: implications of tropomodulin-TM5 or tropomodulin-TM5b complexes in the protofilament and hexagonal organization of membrane skeletons.
The human erythrocyte membrane skeleton consists of hexagonal lattices with junctional complexes containing F-actin protofilaments of approximately 33-37 nm in length. We hypothesize that complexes formed by tropomodulin, a globular capping protein at the pointed end of actin filaments, and tropomyosin (TM), a rod-like molecule of approximately 33-35 nm, may contribute to the formation of protofilaments. We have previously cloned the human tropomodulin complementary DNA and identified human TM isoform 5 (hTM5), a product of the gamma-TM gene, as one of the major TM isoforms in erythrocytes. We now identify TM5b, a product of the alpha-TM gene, to be the second major TM isoform. TM5a, the alternatively spliced isoform of the alpha-TM gene, which differs by 1 exon and has a weaker actin-binding affinity, however, is not present. TM4, encoded by the delta-TM gene, is not present either. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hTM5 comigrated with the slower TM major species in erythrocyte membranes, and hTM5b comigrated with the faster TM major species. TM5b, like TM5, binds strongly to tropomodulin, more so than other TM isoforms. The 2 major TM isoforms, therefore, share several common features: They have 248 residues, are approximately 33-35 nm long, and have high affinities toward F-actin and tropomodulin. These common features may be the key to the mechanism by which protofilaments are formed. Tropomodulin-TM5 or tropomodulin-TM5b complexes may stabilize F-actin in segments of approximately 33-37 nm during erythroid terminal differentiation and may, therefore, function as a molecular ruler. TM5 and TM5b further define the hexagonal geometry of the skeletal network and allow actin-regulatory functions of TMs to be modulated by tropomodulin. (Blood. 2000;95:1473-1480)
Sung
L A
LA
Department of Bioengineering and Center for Molecular Genetics, University of California, San Diego, La Jolla, CA 92093-0412, USA. amysung@bioeng.ucsd.edu
Gao
K M
KM
Yee
L J
LJ
Temm-Grove
C J
CJ
Helfman
D M
DM
Lin
J J
JJ
Mehrpouryan
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000097:blood* / Q000737:chemistry; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000648:ultrastructure; D004910:Erythrocyte Membrane / Q000648:ultrastructure*; D004912:Erythrocytes / Q000378:metabolism*; D015672:Erythroid Precursor Cells / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D008840:Microfilament Proteins / Q000097:blood / Q000737:chemistry; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000097:blood / Q000737:chemistry / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D051348:Tropomodulin; D014335:Tropomyosin / Q000097:blood* / Q000737:chemistry / Q000235:genetics
D002352:Carrier Proteins; D008840:Microfilament Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; C495829:TMOD1 protein, human; C498783:TPM3 protein, human; C495832:Tmod1 protein, rat; C498784:Tpm3 protein, rat; D051348:Tropomodulin; D014335:Tropomyosin