Studies on the membrane-associated proteolytic activities of Escherichia coli

Palmer, S M

Title: Studies on the membrane-associated proteolytic activities of Escherichia coli

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Abstract

The membrane fraction contains three proteolytic activities which can be resolved from whole membrane detergent extracts by DEAE-cellulose chromatography. The first two eluting activities have been previously reported as protease V and protease IV. These two enzymes were further purified by gel permeation HPLC. Protease V has a M. W. of 31,000 in SDS-PAGE gels. Protease IV has a M. W. of 62,000 and exists in two distinct isoforms of pl approx. = 6.7 and 6.9. The third enzyme eluting from the DEAE-cellulose column was further purified by affinity chromatography on Benzamidine-Sepharose 6B. This enzyme, referred to herein as protease VI, is a 43 kdal protein which has not been previously characterized. Protease VI was sensitive to inhibition by the serine protease inhibitors phenylmethylsulfony fluoride (PMSF), diisopropylfluorophosphate (DFP) and p-amino-benzamidine (PAB). Additionally, this enzyme exhibited maximal activity at pH approx. = 8.0. Incubation of whole membrane preparations with (/sup 3/H) DFP resulted in 11 specific proteins acquiring the radioactive label, included in this group of proteins were proteases IV, V, and VI. Several of the DFP-reactive proteins were also shown to bind (/sup 125/I) ampicillin.

Authors:: Palmer, S M

Publication Date:: Wed Jan 01 00:00:00 EST 1986

Research Org.:: Rutgers-the State Univ., New Brunswick, NJ (USA)

OSTI Identifier:: 5281313

Resource Type:: Thesis/Dissertation

Resource Relation:: Other Information: Thesis (Ph. D.)

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; ESCHERICHIA COLI; PROTEOLYSIS; PEPTIDE HYDROLASES; ENZYME ACTIVITY; FRACTIONATION; CELL MEMBRANES; ENZYME INHIBITORS; IODINE 125; LIQUID COLUMN CHROMATOGRAPHY; TRACER TECHNIQUES; TRITIUM COMPOUNDS; BACTERIA; BETA DECAY RADIOISOTOPES; CELL CONSTITUENTS; CHEMICAL REACTIONS; CHROMATOGRAPHY; DAYS LIVING RADIOISOTOPES; DECOMPOSITION; ELECTRON CAPTURE RADIOISOTOPES; ENZYMES; HYDROLASES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; LABELLED COMPOUNDS; MEMBRANES; MICROORGANISMS; NUCLEI; ODD-EVEN NUCLEI; RADIOISOTOPES; SEPARATION PROCESSES; 550201* - Biochemistry- Tracer Techniques

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                    Palmer, S M. Studies on the membrane-associated proteolytic activities of Escherichia coli.  United States: N. p., 1986. 
        Web.

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                    Palmer, S M. Studies on the membrane-associated proteolytic activities of Escherichia coli.  United States.

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                    Palmer, S M. 1986.  
        "Studies on the membrane-associated proteolytic activities of Escherichia coli".  United States.

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@article{osti_5281313,

  title        = {Studies on the membrane-associated proteolytic activities of Escherichia coli},

  author       = {Palmer, S M},

  abstractNote = {The membrane fraction contains three proteolytic activities which can be resolved from whole membrane detergent extracts by DEAE-cellulose chromatography. The first two eluting activities have been previously reported as protease V and protease IV. These two enzymes were further purified by gel permeation HPLC. Protease V has a M. W. of 31,000 in SDS-PAGE gels. Protease IV has a M. W. of 62,000 and exists in two distinct isoforms of pl approx. = 6.7 and 6.9. The third enzyme eluting from the DEAE-cellulose column was further purified by affinity chromatography on Benzamidine-Sepharose 6B. This enzyme, referred to herein as protease VI, is a 43 kdal protein which has not been previously characterized. Protease VI was sensitive to inhibition by the serine protease inhibitors phenylmethylsulfony fluoride (PMSF), diisopropylfluorophosphate (DFP) and p-amino-benzamidine (PAB). Additionally, this enzyme exhibited maximal activity at pH approx. = 8.0. Incubation of whole membrane preparations with (/sup 3/H) DFP resulted in 11 specific proteins acquiring the radioactive label, included in this group of proteins were proteases IV, V, and VI. Several of the DFP-reactive proteins were also shown to bind (/sup 125/I) ampicillin.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/5281313},
  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {Wed Jan 01 00:00:00 EST 1986},

  month        = {Wed Jan 01 00:00:00 EST 1986}

}

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