Recombinant Saccharomyces cerevisiae Ubiquitin carboxyl-terminal hydrolase 4 (DOA4), partial

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Cat. No.

BT44049

Source

Yeast

Synonyms

DOA4; DOS1; MUT4; NPI2; SSV7; UBP4; YDR069C; D4270; YD8554.02C; YD9609.23C; Ubiquitin carboxyl-terminal hydrolase 4; EC 3.4.19.12; Deubiquitinating enzyme 4; Ubiquitin thioesterase 4; Ubiquitin-specific-processing protease 4; Vacuole biogenesis protein SSV7

Purity

>85% (SDS-PAGE)

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Cat. No.

BT44049

Source

E.coli

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT44049

Source

E.coli

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT44049

Source

Baculovirus

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT44049

Source

Mammalian cell

Synonyms

Purity

>85% (SDS-PAGE)

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Description

Definition and Core Functionality

Recombinant Saccharomyces cerevisiae Ubiquitin carboxyl-terminal hydrolase 4 (DOA4) is a truncated or partially expressed version of the yeast deubiquitinating enzyme DOA4. This enzyme catalyzes the removal of ubiquitin from polyubiquitinated substrates, primarily functioning in the ubiquitin-proteasome pathway (UPP) to recycle ubiquitin and facilitate substrate degradation . The "partial" designation indicates that recombinant production may involve truncations or engineered domains to optimize activity or solubility .

Biological Roles

DOA4’s functions span multiple cellular pathways:

Proteasome-Dependent Degradation: Removes ubiquitin from substrates to recycle the molecule and prevent proteasome clogging .
ESCRT-III Regulation: Binds Snf7 (ESCRT-III subunit) to inhibit intralumenal vesicle (ILV) formation, delaying membrane scission until substrate deubiquitination is complete .
Vacuolar Protein Sorting: Collaborates with class E Vps factors (e.g., Vps4) to process ubiquitinated membrane proteins at late endosomes .

Recombinant Production and Applications

Recombinant DOA4 is produced in diverse systems:

Host System	Purity	Applications
E. coli	≥85% (SDS-PAGE)	High-yield expression; enzymatic studies (e.g., ubiquitin chain cleavage)
Pichia pastoris	Secreted form	Cleavage of ubiquitin-fusion proteins (e.g., magainin)
Yeast/Baculovirus	Partial truncations	Structural studies; domain-specific functional analysis (e.g., N-terminal binding)

Key Applications

Ubiquitin Recycling: Critical for maintaining cellular ubiquitin pools during proteasomal degradation .
ESCRT-III Regulation: Overexpression disrupts Snf7 polymerization, inhibiting ILV formation .
In Vitro Assays: Used to study deubiquitination kinetics and substrate specificity .

Product Specs

Form

Lyophilized powder. Note: We will prioritize shipping the format we have in stock. If you have specific format requirements, please note them when ordering, and we will fulfill your request.

Lead Time

Delivery times vary based on purchasing method and location. Please consult your local distributor for specific delivery information. Note: All proteins are shipped with standard blue ice packs. For dry ice shipping, please contact us in advance, as additional charges apply.

Notes

Avoid repeated freezing and thawing. Working aliquots can be stored at 4°C for up to one week.

Reconstitution

Briefly centrifuge the vial before opening to ensure contents are at the bottom. Reconstitute the protein in sterile deionized water to a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our standard final glycerol concentration is 50% for your reference.

Shelf Life

Shelf life depends on several factors, including storage conditions, buffer components, storage temperature, and protein stability. Generally, the liquid form has a shelf life of 6 months at -20°C/-80°C, while the lyophilized form has a shelf life of 12 months at -20°C/-80°C.

Storage Condition

Store at -20°C/-80°C upon receipt. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.

Tag Info

The tag type will be determined during the manufacturing process. If you require a specific tag type, please inform us, and we will prioritize developing it.

Synonyms

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Protein Length

Partial

Purity

>85% (SDS-PAGE)

Species

Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast)

Target Names

DOA4

Uniprot No.

P32571

Target Background

Function

Ubiquitin thioesterase acting at the late endosome/prevacuolar compartment. It recovers ubiquitin from ubiquitinated membrane proteins destined for the vacuole. It also removes ubiquitin from soluble proteins targeted to proteasomes, maintaining free ubiquitin levels. It is involved in ammonium-induced down-regulation of the GAP1 permease, UME3 destruction in response to oxidative stress, and the RAD9 checkpoint response to TOP1 poisons. It also coordinates DNA replication and prevents overreplication.

Gene References Into Functions

1. Doa4 interacts with Snf7 (ESCRT-III) and cleaves Lys63-linked ubiquitin oligomers involved in membrane protein trafficking (PMID: 26427873). 2. Vps20 binding to the Doa4 N-terminus inhibits a non-catalytic Doa4 function that promotes ILV formation (PMID: 23444383). 3. This is the first demonstration of a physiologically relevant DUB (Doa4) subcellular localization signal and illustrates functional diversification of DUB paralogues through evolving alternative spatial signals (PMID: 17145966). 4. Doa4 deubiquitination of cargo proteins and/or MVB sorting machinery components is crucial for proper cargo sorting into the MVB pathway (PMID: 17376168). 5. Dual mechanisms control Doa4-mediated deubiquitination at multivesicular bodies (PMID: 17446860).

Database Links

KEGG: sce:YDR069C

STRING: 4932.YDR069C

Protein Families

Peptidase C19 family

Subcellular Location

Cytoplasm. Late endosome membrane; Peripheral membrane protein. Note=Recruited to the late endosome by BRO1.

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Recombinant Saccharomyces cerevisiae Ubiquitin carboxyl-terminal hydrolase 4 (DOA4), partial

Description

Definition and Core Functionality

Biological Roles

Recombinant Production and Applications

Key Applications

Product Specs

Target Background

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