TY - JOUR
T1 - Effect of Tunicamycin on the Biosynthesis of Human Fibroblast Collagenase
AU - Chua, Chu Chang
AU - Ladda, Roger L.
N1 - Funding Information:
This investigation was supported in part by National Institutes of Health Grant (CA 45056). We thank Dr. A. Eisen for the generous gift of collagenase antibody. The expert technical assistance of Ms. Deborah E. Geiman is grately acknowledged. We thank Drs. V. Bhavanandan, Balvin Chua and George Keller for helpful discussions and Chuty Erves for typing the mansucript.
PY - 1987
Y1 - 1987
N2 - It is generally accepted that collagenase from human fibroblast cultures consists of two proenzymes (Mr 60,000 and 55,000) and two active forms (Mr 50,000 and 43,000). We demonstrated previously that epidermal growth factor (EGF) as well as a number of other growth factors induced the secretion of procollagenase (Mr 60,000, Mr 55,000) into the medium of human fibroblast cultures (Chua et al., 1985). In the presence of tunicamycin and EGF, the secretion of the larger form of procollagenase was suppressed preferentially with concomitant appearance of a new band, Mr 40,000. This Mr 40,000 band could be specifically immunoprecipitated by antibody raised against human collagenase. By two-dimensional peptide mapping, the Mr 40,000 material appeared to have similar composition as the Mr 60,000 band. In a time course study, the Mr 55,000 procollagenase band was the earliest protein to appear in the medium after 1 hour labeling with [35S] -methionine. The Mr 60,000, 50,000 and 43,000 bands appeared after a 2 hour labeling period. Our results indicate that human collagenases are glycosylated proteins and are synthesized via the dolichol phosphate pathway.
AB - It is generally accepted that collagenase from human fibroblast cultures consists of two proenzymes (Mr 60,000 and 55,000) and two active forms (Mr 50,000 and 43,000). We demonstrated previously that epidermal growth factor (EGF) as well as a number of other growth factors induced the secretion of procollagenase (Mr 60,000, Mr 55,000) into the medium of human fibroblast cultures (Chua et al., 1985). In the presence of tunicamycin and EGF, the secretion of the larger form of procollagenase was suppressed preferentially with concomitant appearance of a new band, Mr 40,000. This Mr 40,000 band could be specifically immunoprecipitated by antibody raised against human collagenase. By two-dimensional peptide mapping, the Mr 40,000 material appeared to have similar composition as the Mr 60,000 band. In a time course study, the Mr 55,000 procollagenase band was the earliest protein to appear in the medium after 1 hour labeling with [35S] -methionine. The Mr 60,000, 50,000 and 43,000 bands appeared after a 2 hour labeling period. Our results indicate that human collagenases are glycosylated proteins and are synthesized via the dolichol phosphate pathway.
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U2 - 10.1016/S0174-173X(87)80034-1
DO - 10.1016/S0174-173X(87)80034-1
M3 - Article
C2 - 2822343
AN - SCOPUS:0023270427
VL - 7
SP - 285
EP - 293
JO - Topics in Catalysis
JF - Topics in Catalysis
IS - 4
ER -