A homologue of cathepsin L identified in conditioned medium from Sf9 insect cells

Lindskog, Eva; Svensson, Ingrid; Häggström, Lena

doi:10.1007/s00253-005-0181-9

Cited by 4 publications

(2 citation statements)

References 39 publications

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“…Perhaps the close association of proV-CATH in a complex with CHIA is less prone to premature proteolytic maturation to V-CATH (by either cellular or extracellular proteases) (16,21,22) due to CHIA blocking its prodomain cleavage site, which would not be possible for the chiA-deficient ⌬CH/CA virus.…”

Section: Discussionmentioning

confidence: 99%

Role of Interactions between Autographa californica Multiple Nucleopolyhedrovirus Procathepsin and Chitinase Chitin-Binding or Active-Site Domains in Viral Cathepsin Processing

Hodgson

Arif

Krell

2013

J Virol

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The binding of Autographa californica multiple nucleopolyhedrovirus chitinase (CHIA) to viral cathepsin protease progenitor (proV-CATH) governs cellular/endoplasmic reticulum (ER) coretention of CHIA and proV-CATH, thus coordinating simultaneous cellular release of both host tissue-degrading enzymes upon host cell death. CHIA is a proposed proV-CATH folding chaperone because insertional inactivation of chiA causes production of proV-CATH aggregates that are incompetent for proteolytic maturation into active V-CATH enzyme. We wanted to determine whether the N-terminal chitin-binding domain (CBD, 149 residues) and C-terminal CHIA active-site domain (ASD, 402 residues) of CHIA bind to proV-CATH independently of one another and whether either domain is dispensable for CHIA's putative proV-CATH folding chaperone activity. We demonstrate that N-terminally green fluorescent protein (GFP)-fused CHIA, ASD, and CBD each colocalize with proV-CATH-RFP in ER-like patterns and that both ASD and CBD independently associate with proV-CATH in vivo using bimolecular fluorescence complementation (BiFC) and in vitro using reciprocal nickel-histidine pulldown assays. Altogether, the data from colocalization, BiFC, and reciprocal copurification analyses suggest specific and independent interactions between proV-CATH and both domains of CHIA. These data also demonstrate that either CHIA domain is dispensable for normal proV-CATH processing. Furthermore, in contrast to prior evidence suggesting that a lack of chiA expression causes proV-CATH to become aggregated, insoluble, and unable to mature into V-CATH, a chiA deletion bacmid virus we engineered to express just v-cath produced soluble proV-CATH that was prematurely secreted from cells and proteolytically matured into active V-CATH enzyme.

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Section: Discussionmentioning

confidence: 99%

Role of Interactions between Autographa californica Multiple Nucleopolyhedrovirus Procathepsin and Chitinase Chitin-Binding or Active-Site Domains in Viral Cathepsin Processing

Hodgson

Arif

Krell

2013

J Virol

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“…The proteolytic action may affect amino acid residues essential for enzymatic activity and later degrade the C-terminal portion of FUT9, the region recognized by the anti-FUT9 antibody. Sf9 cells express several classes of proteases, namely cysteine, that are secreted into the extracellular medium (Lindskog et al 2006) and whose activity was shown to increase from the third day of culture onwards (Gotoh et al 2001). The decrease of sFUT9 activity and detection in the supernatant was not due to protein denaturation since we did not observe protein precipitation in the culture medium.…”

Section: Resultsmentioning

confidence: 97%

Stable expression of an active soluble recombinant form of human fucosyltransferase IX in Spodoptera frugiperda Sf9 cells

2007

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A secretory form of human alpha3-fucosyltransferase IX (sFUT9) was overexpressed in Spodoptera frugiperda (Sf9) insect cells using the stable expression vector pIB/V5-His-TOPO and the signal sequence of human interleukin 2 for efficient secretion. sFUT9 was active and its three potential N-glycosylation sites were occupied. sFUT9 efficiently fucosylated the type II acceptors Galbeta4GlcNAC-R and Fucalpha2Galbeta4GlcNAc-R (R = (CH2)3NHCO(CH2)5-NH-biotin) but not the corresponding sialylated acceptor, and only very poorly the type I (Galbeta3GlcNAc-R) related acceptors. sFUT9 showed a clear preference for glycoproteins containing type II acceptors, with values of 121, 113 and 110 microU/million cell for asialofetuin, erythropoietin and asialoerythropoietin, respectively, values approximately 11-fold higher than those obtained for the small acceptors.

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Novel cathepsin L-like protease from dermestid beetle Dermestes frischii maggot

et al. 2011

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A homologue of cathepsin L identified in conditioned medium from Sf9 insect cells

Cited by 4 publications

References 39 publications

Role of Interactions between Autographa californica Multiple Nucleopolyhedrovirus Procathepsin and Chitinase Chitin-Binding or Active-Site Domains in Viral Cathepsin Processing

Role of Interactions between Autographa californica Multiple Nucleopolyhedrovirus Procathepsin and Chitinase Chitin-Binding or Active-Site Domains in Viral Cathepsin Processing

Stable expression of an active soluble recombinant form of human fucosyltransferase IX in Spodoptera frugiperda Sf9 cells

Novel cathepsin L-like protease from dermestid beetle Dermestes frischii maggot

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