Open AccessDetection and subcellular localization of mature protein H, involved in excretion of cloacin DF13
Author(s) -
Oudega Bauke,
Ykema Adrie,
Stegehuis Freek,
Graaf Frits K.
Publication year - 1984
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1984.tb00705.x
Subject(s) - cytoplasm , subcellular localization , escherichia coli , signal peptide , sodium dodecyl sulfate , biochemistry , microbiology and biotechnology , polyacrylamide gel electrophoresis , gel electrophoresis , membrane , chemistry , biology , gene , peptide sequence , enzyme
To investigate whether the pCloDF13 encoded protein H is synthesized as a precursor with a signal sequence and to detect the mature protein H, radioactive labeling experiments with Escherichia coli minicells harboring different pCloDF13 derivatives were carried out. Analysis of the labeled polypeptides on special sodium dodecyl sulfate (SDS)‐polyacrylamide gels revealed that the polypeptide encoded by the pCloDF13 gene H is a precursor with an apparent M r of about 5000. The mature polypeptide had an apparent M r of about 2800. Localization studies revealed that the mature protein H is predominantly located in the membrane fraction, both in the cytoplasmic and in the outer membranes of the minicells.