Literature summary for 7.6.2.2 extracted from

Kodan, A.; Shibata, H.; Matsumoto, T.; Terakado, K.; Sakiyama, K.; Matsuo, M.; Ueda, K.; Kato, H.
Improved expression and purification of human multidrug resistance protein MDR1 from baculovirus-infected insect cells (2009), Protein Expr. Purif., 66, 7-14.

Cloned(Commentary)

Cloned (Comment)	Organism
MDR1, large-scale expression of His10-tagged human MDR1 using a baculovirus/insect expressSF+ cell system	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	-	Homo sapiens	16020	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + H2O + xenobiotic/in	Homo sapiens	multidrug resistance protein MDR1, i.e. P-glycoprotein or ABCB1, is an ATP-dependent efflux pump for various cytotoxic agents, and is implicated in the resistance of human tumors to chemotherapeutic drugs	ADP + phosphate + xenobiotic/out	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His10-tagged MDR1 249fold by metal chelate affinity chromatography and gel filtration, method optimization and evaluation	Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1.7	-	purified recombinant wild-type MDR1, ATPase activity with substrate verapamil	Homo sapiens

Storage Stability

Storage Stability	Organism
most suitable component for maintaining hMDR1 activity is PIPES, rather than citric acid, imidazole, MES, or MOPS. Optimal pH for stabilizing the recombinant enzyme protein during purification is pH 6.5. Addition of CHS preserves 90% of the activity at 25°C after 1 day, whereas little or no activity remains without CHS. The specific ATPase activity is completely diminished after 2 h at 37°C, but 75% of the ATPase activity remains after 24 h at 10°C	Homo sapiens

Organism

most suitable component for maintaining hMDR1 activity is PIPES, rather than citric acid, imidazole, MES, or MOPS. Optimal pH for stabilizing the recombinant enzyme protein during purification is pH 6.5. Addition of CHS preserves 90% of the activity at 25°C after 1 day, whereas little or no activity remains without CHS. The specific ATPase activity is completely diminished after 2 h at 37°C, but 75% of the ATPase activity remains after 24 h at 10°C

Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + H2O + verapamil/in	-	Homo sapiens	ADP + phosphate + verapamil/out	-	?
ATP + H2O + xenobiotic/in	-	Homo sapiens	ADP + phosphate + xenobiotic/out	-	?
ATP + H2O + xenobiotic/in	multidrug resistance protein MDR1, i.e. P-glycoprotein or ABCB1, is an ATP-dependent efflux pump for various cytotoxic agents, and is implicated in the resistance of human tumors to chemotherapeutic drugs	Homo sapiens	ADP + phosphate + xenobiotic/out	-	?

Subunits

Subunits	Comment	Organism
More	three-dimensional structural analysis, overview	Homo sapiens

Synonyms

Synonyms	Comment	Organism
ABCB1	-	Homo sapiens
MDR1	-	Homo sapiens
multidrug resistance protein	-	Homo sapiens
P-glycoprotein	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
10	37	the specific ATPase activity is completely diminished after 2 h at 37°C, but 75% of the ATPase activity remains after 24 h at 10°C	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.4	-	assay at	Homo sapiens

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
6.5	-	optimal pH for stabilizing the recombinant enzyme protein during purification is pH 6.5. hMDR1 remains monodispersed at a pH range of pH 5.5-pH 7.5. In contrast, at a lower pH such as pH 4.5, the protein is precipitated	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens