The R1441C mutation alters the folding properties of the ROC domain of LRRK2.
LRRK2 is a 250 kDa multidomain protein, mutations in which cause familial Parkinson's disease. Previously, we have demonstrated that the R1441C mutation in the ROC domain decreases GTPase activity. Here we show that the R1441C alters the folding properties of the ROC domain, lowering its thermodynamic stability. Similar to small GTPases, binding of different guanosine nucleotides alters the stability of the ROC domain, suggesting that there is an alteration in conformation dependent on GDP or GTP occupying the active site. GTP/GDP bound state also alters the self-interaction of the ROC domain, accentuating the impact of the R1441C mutation on this property. These data suggest a mechanism whereby the R1441C mutation can reduce the GTPase activity of LRRK2, and highlights the possibility of targeting the stability of the ROC domain as a therapeutic avenue in LRRK2 disease.
Li
Yongchao
Y
Department of Biochemistry and Molecular biology, Oklahoma State University, Stillwater, OK 74078, USA.
Dunn
Laura
L
Greggio
Elisa
E
Krumm
Brian
B
Jackson
Graham S
GS
Cookson
Mark R
MR
Lewis
Patrick A
PA
Deng
Junpeng
J
D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D046911:Macromolecular Substances; D009154:Mutation / Q000235:genetics*; D011487:Protein Conformation; D017510:Protein Folding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism
D046911:Macromolecular Substances; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D018631:ras Proteins