Premium
An isoform of actin is associated with the cancer status of ovarian and breast epithelial cells
Author(s) -
Lanner Carita,
Hobbs Hailey,
Gauthier Nicole,
Peltsch Heather,
Cutuli Facundo,
Boyle Amanda,
Naryzhny Stanislav,
Lee Hoyun
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.857.2
Subject(s) - gene isoform , actin , proliferating cell nuclear antigen , ovarian cancer , biology , antibody , microbiology and biotechnology , antigen , immunoprecipitation , breast cancer , chemistry , biochemistry , cancer , cell growth , immunology , gene , genetics
An isoform of actin associated with non‐malignant ovarian and breast epithelial cells was detected in a study investigating cancer specific forms of PCNA (proliferating cell nuclear antigen) in ovarian and breast cancer materials. An antibody (B1 antibody) generated against ten residues from the interdomain connector loop in PCNA, detected not only PCNA, but a novel protein in non‐malignant cells. Immunoprecipitation was performed using the B1 antibody and proteins were cut from gels. In‐gel trypsin digestion was performed for MALDI‐TOF analysis. The novel protein was identified as a form of actin using Mascot peptide fingerprinting software. Immunoblotting with actin antibodies support the identification of the B1 protein as an isoform of actin. A local alignment between PCNA and beta‐actin was performed and found a ten‐residue area of overlap with 50% homology between PCNA and beta‐actin which corresponded to the antigen for the B1 antibody. This homology is likely the basis of the cross reaction with beta‐actin. To avoid the cross reaction with PCNA exhibited by the B1 antibody, a second antibody (ACTB358) was generated to 11 residues starting at residue 358 of beta actin The ACTB358 antibody is highly specific for a novel isoform of actin with a mass of 86 kDa and a pI of 5.5, but does not interact with cytoplasmic or other types of actin. We are investigating the sequence and expression of the novel isoform of actin in non‐malignant and malignant ovarian and breast cells. Support by NOSMFA Research Development Fund