AIM: To comprehensively identify the proteins of tumor relative antigen Ca-Hb3 recognized by colorectal carcinoma monoclonal antibody Hb3. METHODS: Ca-Hb3 was isolated by SDS-polyacrylamide gel electrophoresis (PAGE) followed by digestion with trypsin. Trypsin peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The proteins identified by mass spectrometry were analyzed using bioinformatics. RESULTS: Ca-Hb3 was identified as a CKAP4-like protein by Nano HPLC tandem mass spectrometry analysis. The molecular weight of CKAP4-like protein was 62.02 kDa, including one hydrophobic region, one transmembrane domain, five coiled coils, four glycosylation sites and forty-nine phosphorylation sites. CKAP4-like protein had a high homogeneity with DeltaNp63alpha. The characteristic expression of DeltaNp63alpha that is considered a potential oncogene in the isoforms of p63 was similar to that of Ca-Hb3. CONCLUSION: Ca-Hb3 is probably a CKAP4-like protein, belonging to DeltaNp63alpha isoform of p63 family.
AIM: To comprehensively identify the proteins of tumor relative antigen Ca-Hb3 recognized by colorectal carcinoma monoclonal antibody Hb3. METHODS: Ca-Hb3 was isolated by SDS-polyacrylamide gel electrophoresis (PAGE) followed by digestion with trypsin. Trypsin peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The proteins identified by mass spectrometry were analyzed using bioinformatics. RESULTS: Ca-Hb3 was identified as a CKAP4-like protein by Nano HPLC tandem mass spectrometry analysis. The molecular weight of CKAP4-like protein was 62.02 kDa, including one hydrophobic region, one transmembrane domain, five coiled coils, four glycosylation sites and forty-nine phosphorylation sites. CKAP4-like protein had a high homogeneity with DeltaNp63alpha. The characteristic expression of DeltaNp63alpha that is considered a potential oncogene in the isoforms of p63 was similar to that of Ca-Hb3. CONCLUSION: Ca-Hb3 is probably a CKAP4-like protein, belonging to DeltaNp63alpha isoform of p63 family.
Authors: M Osada; M Ohba; C Kawahara; C Ishioka; R Kanamaru; I Katoh; Y Ikawa; Y Nimura; A Nakagawara; M Obinata; S Ikawa Journal: Nat Med Date: 1998-07 Impact factor: 53.440