Figure 5.
Figure 5. Characterization of human BM stromal cell dipeptidyl peptidases. Stromal cell extracts were separated by ion-exchange chromatography into 2 fractions, E1 and E2, each containing dipeptidyl peptidase activity (A). Dipeptidyl peptidase activity eluted from the column was detected by florigenic assay with 0.1 mM Ala-Pro-AFC, and SDS-PAGE of unboiled samples identified dipeptidyl peptidases of approximately 220 kDa and 170 kDa in E1 and E2, respectively. In contrast, unfractionated stromal extract contained dipeptidyl peptidases of both molecular sizes and RPMI-7951 and CACO-2 cell lines contained only the 170-kDa dipeptidyl peptidase (B). The same samples were immunoprecipitated with mAb F19, M-A261, or a mouse IgG1 isotype control, and the immunoprecipitates were assayed for dipeptidyl peptidase activity (C). Data in panels A-C were representative of 7 independent human stromal cell cultures.

Characterization of human BM stromal cell dipeptidyl peptidases. Stromal cell extracts were separated by ion-exchange chromatography into 2 fractions, E1 and E2, each containing dipeptidyl peptidase activity (A). Dipeptidyl peptidase activity eluted from the column was detected by florigenic assay with 0.1 mM Ala-Pro-AFC, and SDS-PAGE of unboiled samples identified dipeptidyl peptidases of approximately 220 kDa and 170 kDa in E1 and E2, respectively. In contrast, unfractionated stromal extract contained dipeptidyl peptidases of both molecular sizes and RPMI-7951 and CACO-2 cell lines contained only the 170-kDa dipeptidyl peptidase (B). The same samples were immunoprecipitated with mAb F19, M-A261, or a mouse IgG1 isotype control, and the immunoprecipitates were assayed for dipeptidyl peptidase activity (C). Data in panels A-C were representative of 7 independent human stromal cell cultures.

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