An examination of how different mutations at arginine 855 of the androgen receptor result in different androgen insensitivity phenotypes.
Two substitutions at an identical location in the ligand-binding domain (LBD) of the human androgen receptor (AR), R855C and R855H, are associated with complete androgen insensitivity syndrome (AIS) and partial AIS, respectively. Kinetic analysis of the mutant receptors in genital skin fibroblasts and in transfected cells revealed very low total binding (Bmax) and increased rate constants of dissociation (k) for the R855C mutant; and normal Bmax and k, with slightly elevated equilibrium affinity constants (Kd), but decreased transactivational capacity for the R855H mutant. Further analysis of the R855H mutant revealed both thermolability and decreased N/C-terminal inter-actions in the presence and absence of the co-activator transcriptional intermediary factor 2. To establish the nature of these functional differences we have used molecular dynamic modeling to create four-dimensional models of each of the mutant receptors. Molecular dynamic modeling produced profoundly different models for each of the mutants: in modeling of R855C a surprisingly significant distant alteration in the position of helix 12 of the helix 12 positioning of the AR ligand binding domain (AR-LBD) occurs, which would predict severe ligand binding abnormalities and complete AIS; in modeling of R855H, no dramatic effect on the position of helix 12 was seen; thus, binding properties of the receptor are not compromised. Molecular dynamics four-dimensional modeling clearly supports the biochemical and kinetic studies of both mutants. Such novel computational modeling may lead to a better understanding of the structure-function relationships and the molecular mechanics of ligand binding not only of the AR-LBD but also of other nuclear receptors.
Elhaji
Youssef A
YA
Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, 3755 Cote-Ste-Catherine Road, Montreal, Quebec, Canada H3T 1E2.
Wu
Jian Hui
JH
Gottlieb
Bruce
B
Beitel
Lenore K
LK
Alvarado
Carlos
C
Batist
Gerald
G
Trifiro
Mark A
MA
D000328:Adult; D013734:Androgen-Insensitivity Syndrome / Q000235:genetics* / Q000378:metabolism*; D000728:Androgens / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011944:Receptors, Androgen / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature; D015533:Transcriptional Activation
D000728:Androgens; D011944:Receptors, Androgen; D001120:Arginine