TY - JOUR
T1 - Limited in vivo proteolysis of aggregated proteins
AU - Corchero, J. L.
AU - Cubarsí, R.
AU - Enfors, S. o.
AU - Villaverde, A.
PY - 1997/8/18
Y1 - 1997/8/18
N2 - Degradation pathways of insoluble proteins have been analyzed in Escherichia coli by using a N-terminal β-galactosidase fusion protein (VP1LAC) that aggregates immediately after its synthesis. In recombinant E. coli cells, lower molecular mass products, antigenically related to the entire fusion, accumulate together with the entire fusion. In absence of protein synthesis, the insoluble intact protein declines, suggesting that degradation of the recombinant protein also affects aggregated protein. Time course analysis of both soluble and insoluble cell fractions has revealed a limited proteolysis of the insoluble protein that removes the heterologous domain and permits the resulting β-galactosidase fragments to refold and solubilize. Further extensive degradation occurs exclusively on soluble protein. The restricted proteolysis of misfolded, insoluble protein is the initiating event of a subsequent degradative pathway in which rate-limiting steps permit the accumulation of stable degradative intermediates.
AB - Degradation pathways of insoluble proteins have been analyzed in Escherichia coli by using a N-terminal β-galactosidase fusion protein (VP1LAC) that aggregates immediately after its synthesis. In recombinant E. coli cells, lower molecular mass products, antigenically related to the entire fusion, accumulate together with the entire fusion. In absence of protein synthesis, the insoluble intact protein declines, suggesting that degradation of the recombinant protein also affects aggregated protein. Time course analysis of both soluble and insoluble cell fractions has revealed a limited proteolysis of the insoluble protein that removes the heterologous domain and permits the resulting β-galactosidase fragments to refold and solubilize. Further extensive degradation occurs exclusively on soluble protein. The restricted proteolysis of misfolded, insoluble protein is the initiating event of a subsequent degradative pathway in which rate-limiting steps permit the accumulation of stable degradative intermediates.
U2 - https://doi.org/10.1006/bbrc.1997.7132
DO - https://doi.org/10.1006/bbrc.1997.7132
M3 - Article
SN - 0006-291X
VL - 237
SP - 325
EP - 330
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
ER -