RESUMO
NAD+ glycohydrolase activities in serum samples from cancer patients were two- to three-fold higher than the activities in samples from healthy controls. SDS-PAGE analysis of serum samples followed by Western blotting revealed the presence of two proteins of approximately 45 and approximately 21 kDa that were immunoreactive with human CD38-specific monoclonal antibodies T16, HIT2 and OKT 10. These proteins appeared to be more abundant in serum from cancer patients. NAD+ glycohydrolase activity in serum could be enriched by immunoaffinity chromatography by using T16-Sepharose 4B. The results suggest that the relative abundance of proteins immunologically related to CD38 may account for the elevated levels of glycohydrolase activities in serum of tumour patients.
Assuntos
Antígenos CD , Antígenos de Diferenciação/imunologia , Proteínas Sanguíneas/análise , NAD+ Nucleosidase/sangue , NAD+ Nucleosidase/imunologia , Neoplasias/sangue , ADP-Ribosil Ciclase , ADP-Ribosil Ciclase 1 , Adolescente , Adulto , Idoso , Anticorpos Monoclonais/farmacologia , Feminino , Humanos , Masculino , Glicoproteínas de Membrana , Pessoa de Meia-IdadeRESUMO
The penicillin acylase gene (pac) amplified by polymerase chain reaction (PCR) from Escherichia coli (E.coli) ATCC11105 genomic DNA was cloned into pUC9, pEMBL9+ and pCP40 vectors. A penicillin acylase precursor was overexpressed at 26 degrees C from pac-pEMBL9+ and pac-pCP40 constructs transformed into E.coli strains JM109 and pCI857 containing HB101, respectively. With the thermo-inducible pac-pCP40 construct some level of mature alpha and beta subunits and varying degrees of enzyme activity were also detected.
