Nature. 1985 Aug 29-Sep 4;316(6031):817-9. Amplification of - PowerPoint PPT Presentation

Nature. 1985 Aug 29-Sep 4;316(6031):817-9. Amplification of P-glycoprotein genes in multidrug-resistant mammalian cell lines. Riordan JR, Deuchars K, Kartner N, Alon N, Trent J, Ling V.

T HE COMPLEXITY

ABC A1 THE S TRUCTURE INCLUDES SYMMETRY  C OMMON MODULES DEFINE FAMILY FEATURES  E XTRA MODULES DEFINE SPECIFICITIES  Q UATERNARY STRUCTURE IS BASED ON MULTIPLICATION OF SYMMETRY

THE FUNCTION: D YSFUNCTION OF ABC TRANSPORTERS LEADS TO MAJOR HUMAN PATHOLOGIES Out In N N N N NBD NBD NBD NBD NBD NBD NBD NBD C C C C ABCA1 MRP1 CFTR MDR ABCA ABCC ABCB Out N N N N In NBD NBD NBD NBD NBD NBD NBD NBD C C C C C C C C N N N N TAP1/TAP2 ABCG1/ABCG1 ABCG5/ABCG8 ABCB ABCD ABCG

ABC A1 ABC A1 TRANSPORTER Cellular Homeostasis and TANGIER Disease : T HE REVERSE CHOLESTEROL TRANSPORT PARADIGM

ABC A1 AND HDL FORMATION

ABCA1 INFORMS FUNCTIONALLY M F ACTIVATION PROGRAMS  tissue remodelling  i.e. clearance of dead cells  Atheroma progression  i.e. foam cells Brain lesions  resistance to Cerebral Malaria CD3 CD31 F4/ 80 In mice SYTOX MERGE

What is the incidence of ABCA1 on M f reactivity to stimuli ?

ABCA1 positive cells in tissues mAb 1422 anti ABCA1 Extracellular epitope .. are tissue macrophages L. PRADEL

M YELOID CELLS IN THE S PLEEN : HETEROGENEITY A UTOFLUORESCENCE DISGUISED

ABCA1 pos CELLS IN THE SPLEEN ARE R ED P ULP M F AF hi , Lin neg W HOLE SPLEEN ABCA1 S ORTED CELLS : G ENE EXPRESSION BY QRT PCR A CID P HOSPHATASE Relative gene expression TRAP (whole spleen = 10 0 ) PERL ’ S STAINING (FERRIC IRON)

ABCA1 CAN BE EXPRESSED IN YEAST … And goes where it should

ABCA1 GENERATES «LIPID SENSITIVE AND VISIBLE » DOMAINS IN YEAST

2. PS EXPOSURE IS PRESERVED ABCA1 IS FUNCTIONAL IN YEAST  IN YEAST 1. D EAD MUTANT LOSES SURFACE CLUSTERING  IN H E L A 3. LIPID SENSITIVE GROWTH PHENOTYPE  IN YEAST

MEMBRANE L IPID CONTENT AFFECTS ABCA1 - THE CASE OF SL S 50% 100% SL + SM SL 50% - SM

MEMBRANE L IPID CONTENT AFFECTS ABCA1 - THE CASE OF PS  In YEAST cells  In MAMMALIAN cells 100% PS 50% PS + PS - PS

 ABCA1 REQUIRES PS TO FUNCTION LIPID CONTENT ABCA1 ( FRAP ) H OW DOES ABCA1 FUNCTION AFFECT THE MEMBRANE ?

ABC A1  EXPANDS non raft DOMAINS  HAS PROFOUND EFFECTS ON LIPID MICROENVIRONMENT ~ BIOCHEMISTRY , CATIONIC PROBES, FLIM, FRAP, MEMBRANE GUVS ~

FCS PROBE BODIPY SM T HE I NFLUENCE OF ABCA1 Ana Z. Didier M. Tomasz T. ABCA1 A1 MM HeLa

THANKS T O EVERYBODY