Further advances in the production of membrane proteins in Pichia pastoris

Figures & data

Table 1. The 19 high resolution structures of eukaryotic integral membrane proteins produced recombinantly in Pichia pastoris (http://blanco.biomol.uci.edu/mpstruc/listAll/list).

Target	Origin	Ref
G Protein-Coupled Receptors (GPCRs)
A2A adenosine receptor in complex inverse-agonist antibody	Homo sapiens	18
Histamine H1 receptor, complexed with doxepin	Homo sapiens	27
Channels: Potassium and Sodium Ion-Selective
Two-Pore Domain Potassium Channel K2P1.1 (TWIK-1)	Homo sapiens	26
Two-Pore Domain Potassium Channel K2P4.1 (TRAAK)	Homo sapiens	14
Kv1.2 Voltage-gated potassium Channel (full length)	Rattus norvegicus	15
Kv1.2 Voltage-gated potassium Channel	Rattus norvegicus	23
Kv1.2/Kv2.1 Voltage-gated potassium channel chimera	Rattus norvegicus	24
Kv1.2/Kv2.1 Voltage-gated potassium channel chimera	Rattus norvegicus	29
Kir2.2 Inward-Rectifier Potassium Channel (Complete)	Gallus gallus	28
Kir2.2 Inward-Rectifier Potassium Channel in complex with PtdIns(4,5) P 2	Gallus gallus	17
GIRK2 (Kir3.2) G-protein-gated K^+ channel	Mus musculus	31
Channels: Calcium Ion-Selective
Orai Calcium release-activated calcium (CRAC) channel	Drosophila melanogaster	21
Channels: Aquaporins and Glyceroporins
AQP4 aquaporin water channel	Human	19
AQP5 aquaporin water channel (HsAQP5)	Human	20
SoPIP2;1 plant aquaporin (closed conformation)	Spinacia oleracea	30
Aqy1 yeast aquaporin	Pichia pastoris	16
Membrane-Associated Proteins in Eicosanoid and Glutathione Metabolism (MAPEG)
Leukotriene LTC4 Synthase in complex with glutathione	Human	25
ATP Binding Cassette (ABC) Transporters
P-Glycoprotein	Mus musculus	13
P-Glycoprotein	Caenorhabditis elegans	22

Eight new structures (underlined) have been published since the review by Ramón and Marin published in June 2011.¹ Among those, two new protein families are included; GPCRs and ion-selective calcium channels.

Table 2. Common themes in the construct design listed for the eight structures that have been published since the review by Ramón and Marin.

Target		co	ge	sbr	trunc	GFP	T4L	Ref
Human A2A adenosine receptor		yes	yes	-	-	-	-	18
Human Histamine H1 receptor		yes	yes	yes	yes	yes	yes	27
Human Potassium Channel K2P1.1 (TWIK-1)		-	yes	yes	yes	yes	-	26
Human Potassium Channel K2P4.1 (TRAAK)		yes	yes	-	yes	yes	-	14
Chicken Kir2.2 Potassium Channel		-	-	-	-	yes	-	17
Mouse GIRK2 (Kir3.2) G-protein-gated K^+ channel		-	-	-	yes	yes	-	31
Worm P-Glycoprotein		yes	-	-	-	yes	-	22
Drosophila Orai Calcium channel (CRAC)		-	-	yes	yes	yes	-	21

The following shortenings are used; co (codon optimization), ge (N-glycosylation elimination by introducing asn to gln mutations), sbr (disulfide bridge removal), trunc (truncation of hydrophilic termini), GFP (fusion with GFP) and T4L (chimera with T4 lysozyme).

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