The spindle kinesin‐like protein HsEg5 is an autoantigen in systemic lupus erythematosus

Whitehead, Clark M.; Winkfein, Robert J.; Fritzler, Marvin J.; Rattner, J. B.

doi:10.1002/art.1780391005

Cited by 46 publications

(29 citation statements)

References 24 publications

(8 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Therefore, because human autoimmune sera recognize such conserved molecules, they have been used for analysis of cellular proteins (31,32). However, because most autoimmune sera contain antibodies against multiple autoantigens, it is often impossible to determine what component of the autoreactive response results in which cell-staining patterns.…”

Section: Discussionmentioning

confidence: 99%

Dissecting the Cellular Functions of Annexin XI Using Recombinant Human Annexin XI-specific Autoantibodies Cloned by Phage Display

Farnaes

Ditzel

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

Functional studies of cellular proteins are often complicated by the lack of well-defined monoclonal antibodies, the production of which is hampered by the highly conserved nature of these cellular proteins across species. Annexin XI, a member of the Ca 2؉ -dependent, phospholipid-binding protein family, is an example of such a protein and was used in studies to devise a strategy using human autoimmune phage display libraries to generate reagents for biological studies of conserved cellular proteins. An IgG phage display library was generated from bone marrow of an autoimmune patient with high serum antibody titer against annexin XI, which was identified recently as an autoantigen targeted by autoantibodies in several systemic autoimmune diseases. From this phage library, a panel of human monoclonal annexin XI-specific Fabs were isolated and applied to studies of the cellular functions of annexin XI. Confocal microscopy showed a cell cycle-specific redistribution of annexin XI from the cytoplasm to the mitotic spindle. In metaphase, annexin XI was upregulated and costained with ␣-tubulin. The subcellular distribution of annexin XI in COS-7 cells was shown to be Ca 2؉ -dependent, and exhibited a predominantly nuclear pattern at low concentrations and a cytoplasmic pattern at high Ca 2؉ concentrations. Calcyclin, found previously to bind annexin XI in vitro, in vivo coated the nuclear membrane of cultured cell lines and did not colocalize with annexin XI. Ultrastructural analysis by immunoelectron microscopy revealed that annexin XI associated with specific granules in both neutrophils and eosinophils, suggesting a role in the exocytotic pathway. Our results illuminate the multifunctional nature of human annexin XI, provide the first evidence that annexin XI associates with the mitotic spindles and might play a role in cell division, and clearly illustrate the potential of phage display-derived human autoantibodies in broader analyses of the function of highly conserved cellular proteins.

show abstract

Section: Discussionmentioning

confidence: 99%

Dissecting the Cellular Functions of Annexin XI Using Recombinant Human Annexin XI-specific Autoantibodies Cloned by Phage Display

Farnaes

Ditzel

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…28,29 NuMA was detected using a human serum containing autoantibodies to this protein as previously described, at a 1/500 dilution. 47 Acetylated microtubules were detected using a mouse anti-acetylated tubulin antibody (Sigma, T6793) at a 1/1,000 dilution. Gamma-tubulin was was a kind gift from Dr. G. Wahl (Salk Institute for Biological Studies).…”

Section: Methodsmentioning

confidence: 99%

Low-dose laulimalide represents a novel molecular probe for investigating microtubule organization

et al. 2012

View full text Add to dashboard Cite

“…Comparisons with known sequences were performed by BLAST (Altschul et al 1990) on the NCBI Internet Server. The complete 5© sequence of the cDNA was determined by 5© RACE (rapid amplification of cDNA ends) as previously described (Whitehead et al 1996). The 3© end of the cDNA was also determined by RACE using essentially the same protocol except that the dT anchor primer was used rather than oligo(dT) to prime the first-strand cDNA.…”

Section: Methodsmentioning

confidence: 99%

ASE-1: a novel protein of the fibrillar centres of the nucleolus and nucleolus organizer region of mitotic chromosomes

et al. 1997

Self Cite

View full text Add to dashboard Cite

A novel nucleolar component has been identified and cloned using a human autoimmune serum. This antigen, as inferred from the cDNA sequence, is an Mr 55000 protein. Immuno blot analysis, however, of both the native protein and the in vitro translation products of the cDNA showed that they migrate on SDS-PAGE at an apparent molecular mass of 90000 A BLAST search using the cDNA sequence indicated that it is in an antisense orientation to and overlaps the gene of the DNA repair enzyme ERCC-1. An open reading frame, without a translational start site, had been observed by others in this region of the chromosome 19 (19q13.3) and the putative protein was termed ASE-1 (Anti-Sense to ERCC-1). Our cDNA is a full-length equivalent of that open reading frame. ASE-1 was found to contain two domains that are present in a number of nucleolar specific proteins originating from a variety of organisms: a glycine-, arginine- and phenylalanine-rich putative nucleotide interaction domain and an alternating basic/acidic region. Indirect immunofluorescence analysis using antibodies generated to cloned regions of ASE-1 indicated that this protein occurs at the fibrillar centres of the nucleolus in interphase, the putative sites of rDNA transcription, and during cell division it is localized to the nucleolus organizer regions of the chromosomes. ASE-1 co-localises with the RNA polymerase I transcription initiation factor UBF/NOR-90 throughout all stages of the cell cycle and these two proteins associate with each other in vitro.

show abstract

The spindle kinesin‐like protein HsEg5 is an autoantigen in systemic lupus erythematosus

Cited by 46 publications

References 24 publications

Dissecting the Cellular Functions of Annexin XI Using Recombinant Human Annexin XI-specific Autoantibodies Cloned by Phage Display

Dissecting the Cellular Functions of Annexin XI Using Recombinant Human Annexin XI-specific Autoantibodies Cloned by Phage Display

Low-dose laulimalide represents a novel molecular probe for investigating microtubule organization

ASE-1: a novel protein of the fibrillar centres of the nucleolus and nucleolus organizer region of mitotic chromosomes

Contact Info

Product

Resources

About