UTPase or ATPase measurements of puriﬁedABCs have been reported. Only one puriﬁed yeast ABCtransporter, the pleiotropic drug resistance efﬂuxerPdr5p, has been submitted to structural studies.The biochemical study of human ABC transporters isoften more advanced than that of the yeasts. The Pgpprotein responsible for multiple drug resistance (MDR)in human cells is especially well studied. One strongimpetusfor the study ofmammalianABC transportersistheir involvement in diseases. Many mendelian diseasesand complex genetic disorders are caused by ABCtransporters including cystic ﬁbrosis, adrenoleuko-dystrophy, Stargardt disease, Tangier disease, immunedeﬁciencies, progressive familial intrahepatic cholesta-sis, Dublin–Johnson syndrome, Pseudoxanthoma elas-ticum, persistent hyperinsulinemic hypoglycemia of infancy due to focal adenomatous hyperplasia, X-linkedsideroblastosis and anemia, age-related maculardegeneration, familial hypoapoproteinemia, Fundusﬂavimaculatis, Retinitis pigmentosum, cone rod dystro-phy etc. Cell lines isolated from diseased tissues allowmolecular study of the involved ABC transporter.Moreover, a variety of drug-resistant cell lines isavailable from MDR or MDR-related protein (MRP)tissues. Basic studies of human ABC transporters wouldgreatly beneﬁt from heterologous expression of humanABC transporter genes in yeast or other cells, but thistechnology is far from being satisfactory yet. Mean-while,knockouttechnologyin the mouse may be neededto begin to understand the molecular and physiologicalfunctions of the mammalian transporters.
Structure and Biochemical Mechanism
In 1998, the ﬁrst high-resolution structure of a NBD,that from the histidine ABC importer HisP, wasreported. Five years later, about six related structureswere available and a consensus view emerged. NBDs areorganized as dimers and two molecules of ATP arebound at their interface. Each nucleotide-binding sitecomprises a Walker A motiffrom monomer 1 and the Cmotif from monomer 2. This results from a “head-to-tail” arrangement of the two interacting monomers.This is supported by biochemical arguments and iscoherent with the cooperative hydrolysis for ATPhydrolysis observed with MalK.More recently, three structures of complete dimericABC transporters comprising both NDB and TMD wereobtained: that of the presumed phospholipid ﬂippaseMsbA from
and that of thevitamin B12 importer BtuCD from
. Thestructures obtained were dissimilar, which may not betoo surprising taking into account the different con-ditions used, the different numbers of TM spans and thedifferent functions (import or export) of the proteinsanalyzed. No generalization can be made, for instance,on the angle between the TM spans and the membraneplane or on the identiﬁcation of the interaction domainsbetween the TM spans. The nature of communicationbetween the NBD and TMD is variable and carried outeither through the long and complex so-called intra-cellulardomainnamed ICDinMsbA,orthroughashort
-shapedlinkerbetweenthetransmembranespans6and7inBtuCD.Thenature,thesize,theorientation,andthelocation of the so-called chamber (or water channel, orpore, or cone) presumed to be involved in substratebinding are also variable. No consensus interactionpoints between the NBDs (open orclosed conformation)were observed. Obviously, more structures are neededon several transporters carrying out similar functions,such as drug efﬂux, for instance, to clarify these issuesand to reach a consensus interpretation of the basicstructural elements involved in the transport and in thecoupling mechanism.In contrast, recent analyses at the electronmicroscopy level of a bacteria (BmrA or YccV from
) and a yeast (Pdr5p) drug efﬂux ABCtransporter came to a remarkably coherent set of conclusions. In both cases, the basic structural unitseems to comprise four joining NBDs (that correspondsto two full-size Pdr5p or four half-size BmrA), which are
Example of proteins associated to bacterial and mammalian ABC transporters.
OutMembraneInNBD1NBD2Periplasmic-bindingreceptorSubstrateTransmembrane domainCytoplasmic nucleotide-binding domainOutNInN-terminalextensionPoresubunitNCCNBD2NBD1