PMID: 9435084Jan 22, 1998Paper

Impaired secretion of a hydrophobic cutinase by Saccharomyces cerevisiae correlates with an increased association with immunoglobulin heavy-chain binding protein (BiP)

Applied and Environmental Microbiology
C M SagtC T Verrips

Abstract

This study focuses on the different efficiencies of secretion of two fungal cutinases by Saccharomyces cerevisiae, a wild-type cutinase (CY000) and a hydrophobic mutant cutinase (CY028). Both cutinases are placed under control of the GAL7 promoter, by which the expression levels can be regulated. Wild-type cutinase was secreted at up to 25 mg per g (dry weight), while CY028 was secreted at a level of 2 mg per g (dry weight); this difference is nearly independent of the expression level. Pulse-chase experiments revealed that whereas CY000 cutinase is secreted, CY028 is irreversibly retained in the cell. Immunogold labelling followed by electron microscopy revealed colocalization of CY028 with immunoglobulin heavy-chain binding protein (BiP) in the endoplasmic reticulum (ER). The increase of wild-type cutinase expression did not result in higher levels of the molecular chaperone BiP, but BiP levels are raised by increased induction of the hydrophobic mutant cutinase. Immunoprecipitation studies showed that in contrast to the wild-type cutinase, the hydrophobic mutant cutinase interacts with BiP. These results indicate that the introduction of two exposed hydrophobic patches in cutinase results in a higher affinity for BiP which m...Continue Reading

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Citations

Oct 31, 2000·Applied and Environmental Microbiology·C M SagtC T Verrips
Feb 1, 2011·Applied and Environmental Microbiology·Ryo TakemasaTadayuki Imanaka
Sep 23, 2003·Protein Science : a Publication of the Protein Society·Stefania BroccaMarina Lotti
Sep 4, 2004·Applied and Environmental Microbiology·Marta LombrañaJuan F Martín
May 7, 2002·Biotechnology and Bioengineering·Cecília R C CaladoLuis P Fonseca
Jun 13, 2000·Protein Science : a Publication of the Protein Society·S BroccaM Lotti
Jan 11, 2005·Applied and Environmental Microbiology·Edward DolkTheo Verrips
Aug 10, 1999·Biotechnology and Bioengineering·A L Santerre HenriksenJ Nielsen
Sep 25, 2019·Protein Expression and Purification·Nan WangDeHu Liu
Aug 4, 1998·Applied and Environmental Microbiology·I A van GemerenC T Verrips
Apr 13, 2020·Progress in Retinal and Eye Research·Luminita ParaoanIan Grierson
Aug 10, 2001·Fungal Genetics and Biology : FG & B·A ConesaC A van den Hondel
Jun 23, 2000·Enzyme and Microbial Technology·T VerripsabC Sagt

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