Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells

A pool of 10 calmodulin-binding proteins (CBPs) was isolated from Chinese hamster ovary (CHO) cells via calmodulin (CaM)-Sepharose affinity chromatography. One of these ten isolated CBPs with a molecular mass of 52 kD was also found to be present in isolated CHO cell mitotic spindles. Affinity-purif...

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Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1986
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Articles
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114387/
https://www.ncbi.nlm.nih.gov/pubmed/3536955
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collection PubMed
description A pool of 10 calmodulin-binding proteins (CBPs) was isolated from Chinese hamster ovary (CHO) cells via calmodulin (CaM)-Sepharose affinity chromatography. One of these ten isolated CBPs with a molecular mass of 52 kD was also found to be present in isolated CHO cell mitotic spindles. Affinity-purified antibodies generated against this pool of isolated CBPs recognize a single 52-kD protein in isolated CHO cell mitotic spindles by immunoblot analysis. Immunofluorescence examination of CHO, 3T3, NRK, PTK-2, and HeLa cells resulted in a distinct pattern of mitotic spindle fluorescence. The localization pattern of this 52-kD CBP directly parallels that of CaM in the spindle apparatus throughout the various stages of mitosis. Interestingly, there was no association of this 52-kD CBP with cytoplasmic microtubules. As is the case with CaM, the localization pattern of the 52-kD CBP in interphase cells is diffuse within the cytoplasm and is not associated with any discrete, cellular structures. This 52-kD CBP appears to represent the first mitotic spindle-specific calmodulin- binding protein identified and represents an initial step toward the ultimate determination of CaM function in the mitotic spindle apparatus.
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spelling pubmed-21143872008-05-01 Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells J Cell Biol Articles A pool of 10 calmodulin-binding proteins (CBPs) was isolated from Chinese hamster ovary (CHO) cells via calmodulin (CaM)-Sepharose affinity chromatography. One of these ten isolated CBPs with a molecular mass of 52 kD was also found to be present in isolated CHO cell mitotic spindles. Affinity-purified antibodies generated against this pool of isolated CBPs recognize a single 52-kD protein in isolated CHO cell mitotic spindles by immunoblot analysis. Immunofluorescence examination of CHO, 3T3, NRK, PTK-2, and HeLa cells resulted in a distinct pattern of mitotic spindle fluorescence. The localization pattern of this 52-kD CBP directly parallels that of CaM in the spindle apparatus throughout the various stages of mitosis. Interestingly, there was no association of this 52-kD CBP with cytoplasmic microtubules. As is the case with CaM, the localization pattern of the 52-kD CBP in interphase cells is diffuse within the cytoplasm and is not associated with any discrete, cellular structures. This 52-kD CBP appears to represent the first mitotic spindle-specific calmodulin- binding protein identified and represents an initial step toward the ultimate determination of CaM function in the mitotic spindle apparatus. The Rockefeller University Press 1986-11-01 /pmc/articles/PMC2114387/ /pubmed/3536955 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title_full Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title_fullStr Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title_full_unstemmed Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title_short Identification of a 52-kD calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
title_sort identification of a 52-kd calmodulin-binding protein associated with the mitotic spindle apparatus in mammalian cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2114387/
https://www.ncbi.nlm.nih.gov/pubmed/3536955

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