Effects of two familial hypertrophic cardiomyopathy-causing mutations on alpha-tropomyosin structure and function.
Missense mutations in alpha-tropomyosin can cause familial hypertrophic cardiomyopathy. The effects of two of these, Asp175Asn and Glu180Gly, have been tested on the structure and function of recombinant human tropomyosin expressed in Escherichia coli. The F-actin affinity (measured by cosedimentation) of Glu180Gly was similar to that of wild-type, but Asp175Asn was more than 2-fold weaker, whether or not troponin was present. The mutations had no apparent effect on the affinity of tropomyosin for troponin. The mutations had a small effect on the overall stability (measured using circular dichroism) but caused increased local flexibility or decreased local stability, as evaluated by the higher excimer/monomer ratios of tropomyosin labeled with pyrene maleimide at Cys 190. The pyrene-labeled tropomyosins differed in their response to myosin S1 binding to the actin-tropomyosin filament. The conformations of the two mutants were different from each other and from wild-type in the myosin S1-induced on-state of the thin filament. Even though both mutant tropomyosins bound cooperatively to actin, they did not respond with the same conformational change as wild-type when myosin S1 switched the thin filament from the off- to the on-state.
Golitsina
N
N
Boston Biomedical Research Institute, Massachusetts 02114, USA.
An
Y
Y
Greenfield
N J
NJ
Thierfelder
L
L
Iizuka
K
K
Seidman
J G
JG
Seidman
C E
CE
Lehrer
S S
SS
Hitchcock-DeGregori
S E
SE
D000199:Actins / Q000378:metabolism / Q000494:pharmacology; D002312:Cardiomyopathy, Hypertrophic / Q000209:etiology* / Q000235:genetics* / Q000378:metabolism; D004347:Drug Interactions; D006801:Humans; D009154:Mutation; D015879:Myosin Subfragments / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding / Q000235:genetics; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014335:Tropomyosin / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D014336:Troponin / Q000378:metabolism
D000199:Actins; D015879:Myosin Subfragments; D014335:Tropomyosin; D014336:Troponin