Identification of a 34-kD polypeptide as a light chain of microtubule-associated protein-1 (MAP-1) and its association with a MAP-1 peptide that binds to microtubules (original) (raw)
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Characterization of a new 120 kDa microtubule-associated protein (MAP) of rat brain
Neuroscience Letters, 1991
A novel protein was identified in rat brain microtubules using a monoclonal antibody. The immunoreactive protein is a microtubule-associated protein (MAP) by the criteria of co-purification with tubulin through repeated cycles of microtubule polymerisation in vitro. It belongs to the class of thermostable MAPs and runs as a closely spaced polypeptide doublet of 120 kDa on SDS PAGE gels. MAP-120 kDa is brain-and neuron-specific and is localized predominantly in Purkinje cell bodies and dendrites in the cerebellum and in dendritic compartments of pyramidal and granule neurons in the hippocampus and dentate gyrus. During postnatal brain development, MAP-120 kD levels increase about 3 to 4-fold.
2002
The microtubule-associated proteins 1A (MAP1A) and 1B (MAP1B) are distantly related protein complexes consisting of heavy and light chains and are thought to play a role in regulating the neuronal cytoskeleton, MAP1B during neuritogenesis and MAP1A in mature neurons. To elucidate functional differences between MAP1B and MAP1A and to determine the role of the light chain in the MAP1A protein complex, we chose to investigate the functional properties of the light chain of MAP1A (LC2) and compare them with the light chain of MAP1B (LC1). We found that LC2 binds to microtubules in vivo and in vitro and induces rapid polymerization of tubulin. A microtubule-binding domain in its NH 2 terminus was found to be necessary and sufficient for these activities. The analysis of LC1 revealed that it too bound to microtubules and induced tubulin polymeriza-tion via a crucial but structurally unrelated NH 2 -terminal domain. The two light chains differed, however, in their effects on microtubule bundling and stability in vivo. Furthermore, we identified actin filament binding domains located at the COOH terminus of LC2 and LC1 and obtained evidence that binding to actin filaments is attributable to direct interaction with actin. Our findings establish LC2 as a crucial determinant of MAP1A function, reveal LC2 as a potential linker of neuronal microtubules and microfilaments, and suggest that the postnatal substitution of MAP1B by MAP1A leads to expression of a protein with an overlapping but distinct set of functions.
Journal of Biological Chemistry, 2004
The related high molecular mass microtubuleassociated proteins (MAPs) MAP1A and MAP1B are predominantly expressed in the nervous system and are involved in axon guidance and synaptic function. MAP1B is implicated in fragile X mental retardation, giant axonal neuropathy, and ataxia type 1. We report the functional characterization of a novel member of the microtubule-associated protein 1 family, which we termed MAP1S (corresponding to sequence data bank entries for VCY2IP1 and C19ORF5). MAP1S contains the three hallmark domains of the microtubuleassociated protein 1 family but hardly any additional sequences. It decorates neuronal microtubules and copurifies with tubulin from brain. MAP1S is synthesized as a precursor protein that is partially cleaved into heavy and light chains in a tissue-specific manner. Heavy and light chains interact to form the MAP1S complex. The light chain binds, bundles, and stabilizes microtubules and binds to actin. The heavy chain appears to regulate light chain activity. In contrast to MAP1A and MAP1B, MAP1S is expressed in a wide range of tissues in addition to neurons and represents the non-neuronal counterpart of this cytolinker family.
Identification of a novel microtubule-binding domain in microtubule-associated protein 1A (MAP1A)
1994
Several microtubule-associated proteins (MAPs) have been shown to bind to microtubules via short sequences with repeated amino acids motifs. A microtubule-binding domain has hitherto not been defined for the adult brain microtubule-associated protein 1A (MAP1A). We have searched for a microtubule-binding domain by expressing different protein regions of MAP1A in cultured cell lines using cDNA constructs. One construct included an area with homology to the microtubule-binding domain of MAP1B (Noble et al. (1989) J. Cell Biol. 109, 437-448), but this did not bind to microtubules in transfected cells. Further investigation of other areas of MAP1A revealed a protein domain, capable of autonomously binding to microtubules, which bears no homology to any previously described microtubule-binding sequence. This MAP1A domain is rich in charged amino acids, as are other mammalian microtubule-binding domains, but unlike them has no identifiable sequence repeats. Whereas all previously describe...
Identification of a Neural Cell Specific Variant of Microtubule-Associated Protein 4
Cell Structure and Function, 2005
The microtubule-binding domain of MAP4, a ubiquitous microtubule-associated protein, contains a region rich in proline and basic residues (proline-rich region). We searched the bovine adrenal gland for MAP4 isoforms, and identified a novel variant lacking 72 consecutive amino acid residues within the proline-rich region, as compared with the full-length MAP4. The amino acid sequence of the missing region was highly conserved (about 85% identity/similarity) among the corresponding regions of bovine, human, mouse, and rat MAP4, which suggested the functional significance of this region. A comparison of the genomic sequence with the cDNA sequence revealed that the missing region is encoded by a single exon. A MAP4 variant cDNA homologous to the bovine form was also detected in rat cells, suggesting that the new variant can be generated by alternative splicing, not only in bovine but also in other mammalian species. The mRNA expression of the novel isoform was restricted to the brain and the adrenal medulla, suggesting that this isoform is specific to a certain cell type. Using a bacterially expressed fragment corresponding to the microtubule-binding domain of the novel isoform, we analyzed its in vitro characteristics. The fragment induced microtubule assembly and bound to preformed microtubules, but the activities were slightly lower than those of the conventional MAP4 fragment, which carries the fulllength proline-rich region. The microtubules assembled in the presence of the fragment failed to be bundled. Instead, a constant spacing between neighboring microtubules was observed.
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 2006
The microtubule-associated proteins MAP1A and MAP1B are related but distinct multi-subunit protein complexes that consist of heavy and light chains. The predominant forms of these complexes are homotypic, i.e. they consist of a MAP1A heavy chain associated with MAP1A light chains or a MAP1B heavy chain associated with MAP1B light chains, respectively. In addition, MAP1A and MAP1B can exchange subunits and form heterotypic complexes consisting of a MAP1A heavy chain associated with MAP1B light chains which might play a role in a transition period of neuronal differentiation. Here we extend previous findings by confirming that heterotypic MAP1B heavy chain-MAP1A light chain complexes also exist in the developing murine brain. We show that these complexes form through interaction of homologous domains conserved in heavy and light chains of MAP1A and MAP1B. Likewise, conserved domains of the MAP1A and MAP1B light chains account for formation of light chain heterodimers. By yeast 2-hybrid analysis we located the light chain binding domain on the heavy chain to amino acids 211-508, thereby defining a new functional subdomain.
Cell Biology International Reports, 1986
A monoclonal antibody to neuronal microtubule-associated protein MAP-2 was produced, Immunoblotting of lysates of cultured cells revealed that the antibody, called MA-01, bound to a protein of M 210 kDa. Double immunofluorescence microscopy showed Fhat the antibody stained microtubules. No fibrillar structures were observed in cells treated with Colcemid, but the antibody stained vinblastlne paracrystals, In cytochalasin B-treated Leydig cells, MA-01 antibody stained star-like structures that codistributed with actin patches and with a star-like arrangement of vimentin, These observations indicate that the protein immunologically related to MAP-2 in Leydig cells could be involved in the interaction of microtubules with intermediate filaments or microf ilaments.
Heterogeneity of microtubule-associated protein (MAP2) in vertebrate brains
Brain Research, 1987
We have utilized monoclonal anti ies to investigate the antigenic diversity of MAIT-immunoreactive proteins in the nervous system of vertebrates. We found that domains defined by the monocional antibodies differed in their conservation across vertebrate evolution, ranging from wide cross-reactivity with almost all vertebrates (mammals, birds, reptiles and amphibians) to a very limited cross-reactivity with only few mammalian *es. Wowever. we did not find MAP?-immunoreactive proteins in fish species with either of the monoclonal or polyclonal anti. There was also a significant divergence in the apparent molecular weight of MAPZ, even in closely related species. For example. rent species of wild mice and strains of laboratory mice showed variations of up to 30 kDa in their apparent molecular mass. Using alkaline phospkata. under conditions that dephosphorylate neurofilaments. we showed that the observed heterogeneity was not the result of variations in the phosphate content. The heterogeneity in molecular weight of MAP2 may, therefore, be the result of changes in primary structure, transcriptional variations or different post-translational modifications. The heterogeneity of MAP2, as well as its specific distribution and implicated interactions with other molecules. underscore the complexity of MAP2 andtential for structural and function rsity. The phylogenic analysis of such a complex molecule also provides a method to est the uniqueness of monoclonal an ies and the degree of their conservation for their corresponding epitopes. 19.23.39.44 AP2 can be cleaved into two structural domains: a fragment which binds the microtubules and promotes tubulin assembly':'and the arm projecting from the microtubule surface which contains a binding site for the regulatory subunit of CA
Experimental Cell Research, 1985
We have obtained several hybridoma clones producing antibodies to microtubule-associated proteins (MAPs) from bovine brain. Interaction of one of these antibodies, named RN 17, with cultured cells was studied by indirect immunofluorescence and immunoelectron microscopy. RN 17 antibody recognized both high molecular weight (HMW) MAPs, MAP 1 and MAP 2, in immunoblotting reaction with brain microtubules. In lysates of cultured cells, it bound to a protein doublet with a molecular weight of 100 kD. By immunofluorescence microscopy we showed that RN 17 antibody stained cytoplasmic fibrils, mitotic spindles and small particles in the cytoplasm of various cultured cells. The cytoplasmic fibrils were identified as both microtubules and intermediate filaments by double fluorescence microscopy and by their response to colcemid and 0.6 M KCl. This identification was confirmed by immunoelectron microscopy which also showed that the particles stained by RN 17 antibody are coated vesicles. Thus, cultured non-neural cells may contain a novel protein that binds to microtubules, intermediate filaments, and coated vesicles.