Linked Life Data

15979569

Source:http://linkedlifedata.com/resource/pubmed/id/15979569

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2005-7-4
pubmed:abstractText
The Kunitz type protease inhibitor aprotinin, containing three intramolecular disulfide bonds, was expressed on the surface of Escherichia coli by Autodisplay. For this purpose, the aprotinin gene was fused in-frame to the transporter domain encoding DNA region of the AIDA-I autotransporter protein. Culture of cells supplied with the artificial gene at reducing conditions resulted in the translocation of aprotinin to the cell surface. Correct folding of aprotinin was shown by high affinity to its target enzyme HLE. No surface translocation was detectable under non-reducing conditions, indicating the degradation of aprotinin in the periplasm. By the use of periplasmic-protease defective E. coli strains PW147, PW151, and PW152, under non-reducing conditions, significant amounts of aprotinin appeared in the periplasm but not at the surface. Our results indicate that aprotinin molecules, reaching stable conformation before transport across the outer membrane, are degraded in the periplasm due to proteolysis. In case folding can be prevented, i.e., by blocking disulfide bond formation in the periplasm, aprotinin is translocated and can adopt its active conformation at the cell surface.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0006-291X
pubmed:author
pubmed:issnType
Print
pubmed:day
12
pubmed:volume
333
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1218-26
pubmed:meshHeading
pubmed:year
2005
pubmed:articleTitle
Autodisplay of the protease inhibitor aprotinin in Escherichia coli.
pubmed:affiliation
Bioanalytics, Institute for Pharmaceutical and Medicinal Chemistry, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, D-40225 Düsseldorf, Germany. joachim.jose@uni-duesseldorf.de
pubmed:publicationType
Journal Article