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Cellular distribution of activity for three enzymes with maltose binding protein as fusion partner and the structural implications

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dc.contributor.authorSARIYAR AKBULUT, BERNA
dc.contributor.authorsUtkur, Ozde F.; Akbulut, Berna Sariyar; Hortacsu, Amable
dc.date.accessioned2022-03-12T17:47:56Z
dc.date.available2022-03-12T17:47:56Z
dc.date.issued2010
dc.description.abstractThe bacterial SEC pathway is commonly used for secretion of heterologous proteins in E. coli by fusing them to transported proteins to facilitate downstream processing. While some proteins are translocated very efficiently, some reside in the cytoplasm. In this work, maltose binding protein (MBP) was fused to 3 cytoplamic enzymes from Thermus thermophilus (serine protease, 251 residues; glucose isomerase, 381 residues; pullulanase, 718 residues) to study the protein transport from the cytoplasm by quantifying the distribution of activities in different cellular compartments. Pullulanase activity was harvested exclusively in the periplasm; however, glucose isomerase activity was harvested exclusively in the cytoplasm. Considerable serine protease activity was found in the periplasm, but after 10 h of induction activity dropped sharply and no activity was found thereafter in either compartment. This was attributed to the instability of the plasmid probably caused by the proteolytic activity of the protease Computations of hypothetical folding rates and secondary structure contents of the proteins showed that folding rates, in addition to alpha-helix and beta-sheet contents of proteins, could be important determinants for efficient translocation by the SEC pathway. These results may give clues to predict whether a protein would be a suitable fusion tail for periplasmic transport with MBP.
dc.identifier.doi10.3906/kim-0901-37
dc.identifier.issn1300-0527
dc.identifier.urihttps://hdl.handle.net/11424/229865
dc.identifier.wosWOS:000274901300001
dc.language.isoeng
dc.publisherSCIENTIFIC TECHNICAL RESEARCH COUNCIL TURKEY-TUBITAK
dc.relation.ispartofTURKISH JOURNAL OF CHEMISTRY
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectMBP fusion protein
dc.subjectperiplasmic secretion
dc.subjectprotein length
dc.subjectsecondary structure
dc.subjectfolding rate
dc.subjectAMINO-ACID-SEQUENCE
dc.subjectESCHERICHIA-COLI
dc.subjectFOLDING RATES
dc.subjectBETA-HAIRPIN
dc.subjectGLUCOSE-ISOMERASE
dc.subjectPREDICTION
dc.subjectTRANSLOCATION
dc.subjectEXPRESSION
dc.subjectMECHANISM
dc.subjectDYNAMICS
dc.titleCellular distribution of activity for three enzymes with maltose binding protein as fusion partner and the structural implications
dc.typearticle
dspace.entity.typePublication
local.avesis.id6fa7e573-20a9-4c70-b9de-a0939eeed3b9
local.import.packageSS17
local.indexed.atWOS
local.indexed.atSCOPUS
local.indexed.atTRDIZIN
local.journal.numberofpages13
oaire.citation.endPage13
oaire.citation.issue1
oaire.citation.startPage1
oaire.citation.titleTURKISH JOURNAL OF CHEMISTRY
oaire.citation.volume34
relation.isAuthorOfPublicationa9f127d3-8332-44dd-a532-34f3ef20bdb5
relation.isAuthorOfPublication.latestForDiscoverya9f127d3-8332-44dd-a532-34f3ef20bdb5

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