Simultaneous red/green dual fluorescence detection on electroblots using BODIPY TR-X succinimidyl ester and ELF 39 phosphate.
AuthorsMartin K, Hart C, Schulenberg B, Jones L, Patton WF
JournalProteomics
PubMed ID11987124
'A two-color fluorescence detection method is described based upon covalently coupling the succinimidyl ester of BODIPY TR-X dye to proteins immobilized on polyvinylidene difluoride membranes, followed by detection of target proteins using the fluorogenic, precipitating substrate ELF 39-phosphate in combination with alkaline phosphatase conjugated reporter molecules. This results in all ... More
Cdc42-interacting protein 4 mediates binding of the Wiskott-Aldrich syndrome protein to microtubules.
AuthorsTian L, Nelson DL, Stewart DM
JournalJ Biol Chem
PubMed ID10713100
'The Wiskott-Aldrich syndrome is an inherited X-linked immunodeficiency characterized by thrombocytopenia, eczema, and a tendency toward lymphoid malignancy. Lymphocytes from affected individuals have cytoskeletal abnormalities, and monocytes show impaired motility. The Wiskott-Aldrich syndrome protein (WASP) is a multi-domain protein involved in cytoskeletal organization. In a two-hybrid screen, we identified the ... More
Abnormal spindle protein, Asp, and the integrity of mitotic centrosomal microtubule organizing centers.
Authorsdo Carmo Avides M, Glover DM
JournalScience
PubMed ID10073938
The product of the abnormal spindle (asp) gene was found to be an asymmetrically localized component of the centrosome during mitosis, required to focus the poles of the mitotic spindle in vivo. Removing Asp protein function from Drosophila melanogaster embryo extracts, either by mutation or immunodepletion, resulted in loss of ... More
Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons.
AuthorsFrancis F, Koulakoff A, Boucher D, Chafey P, Schaar B, Vinet MC, Friocourt G, McDonnell N, Reiner O, Kahn A, McConnell SK, Berwald-Netter Y, Denoulet P, Chelly J
JournalNeuron
PubMed ID10399932
Recently, we and others reported that the doublecortin gene is responsible for X-linked lissencephaly and subcortical laminar heterotopia. Here, we show that Doublecortin is expressed in the brain throughout the period of corticogenesis in migrating and differentiating neurons. Immunohistochemical studies show its localization in the soma and leading processes of ... More
Purification and characterization of native conventional kinesin, HSET, and CENP-E from mitotic hela cells.
AuthorsDeLuca JG, Newton CN, Himes RH, Jordan MA, Wilson L
JournalJ Biol Chem
PubMed ID11382767
We have developed a strategy for the purification of native microtubule motor proteins from mitotic HeLa cells and describe here the purification and characterization of human conventional kinesin and two human kinesin-related proteins, HSET and CENP-E. We found that the 120-kDa HeLa cell conventional kinesin is an active motor that ... More
The plane of first cleavage is not related to the distribution of sperm components in the mouse.
AuthorsDavies TJ, Gardner RL
JournalHum Reprod
PubMed ID12202427
BACKGROUND: Marking experiments using phytohaemagglutinin (PHA) and Concanavalin A (ConA) have suggested that the first cleavage plane is related to the point of sperm entry. Because of concerns about the specificity of lectin binding, the distribution of sperm components has been investigated directly. METHODS AND RESULTS: The sperm tail could ... More
The cell cycle-dependent localization of the CP190 centrosomal protein is determined by the coordinate action of two separable domains.
AuthorsOegema K, Whitfield WG, Alberts B
JournalJ Cell Biol
PubMed ID8522588
CP190, a protein of 1,096 amino acids from Drosophila melanogaster, oscillates in a cell cycle-specific manner between the nucleus during interphase, and the centrosome during mitosis. To characterize the regions of CP190 responsible for its dynamic behavior, we injected rhodamine-labeled fusion proteins spanning most of CP190 into early Drosophila embryos, ... More