Biomolecular interactions between human recombinant beta-MyHC and cMyBP-Cs implicated in familial hypertrophic cardiomyopathy.
OBJECTIVE
Cardiac myosin-binding protein C (cMyBP-C) is a component of sarcomere that contains at least three putative myosin-binding sites. Mutations in its gene are implicated in familial hypertrophic cardiomyopathy (FHC) and most of them are predicted to produce C-terminal truncated cMyBP-Cs. The aim of the present study was to analyze whether cMyBP-C truncated mutants resulting from FHC mutations interact in vitro with human beta-MyHC.


METHODS
Recombinant proteins were produced using the baculovirus/insect cell system, and wild type and three truncated cMyBP-Cs were purified using metal affinity chromatography. The interaction between recombinant proteins was analyzed in real time using biosensor technology on immobilized anti-beta-MyHC antibodies.


RESULTS
Biomolecular interaction with beta-MyHC was detected for both wild type cMyBP-C and a truncated mutant lacking half of the C-terminal C10 domain. In contrast, no interaction with beta-MyHC was found for two truncated cMyBP-Cs lacking at least the C5-C9 region.


CONCLUSIONS
Biosensor technology allows in vitro analysis of the interaction between human beta-MyHC and cMyBP-C mutants resulting from FHC mutations. The data show that the interaction depends on the size of the truncation. This suggests that, in the context of FHC, impairment of suitable interaction between beta-MyHC and some of the truncated cMyBP-Cs may promote degradation of the truncated proteins and therefore contribute to the development of the disease.
Flavigny
Jeanne
J
INSERM U582, Institut de Myologie, Bâtiment Babinski, CHU Pitié-Salpêtrière, 47 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Robert
Philippe
P
Camelin
Jean-Claude
JC
Schwartz
Ketty
K
Carrier
Lucie
L
Berrebi-Bertrand
Isabelle
I
D000818:Animals; D016367:Baculoviridae; D019149:Bioreactors; D015374:Biosensing Techniques; D024741:Cardiomyopathy, Hypertrophic, Familial / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009154:Mutation; D018995:Myosin Heavy Chains / Q000235:genetics* / Q000378:metabolism; D024501:Nonmuscle Myosin Type IIB; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera; D020349:Surface Plasmon Resonance; D000099163:Myosin Binding Protein C
D002352:Carrier Proteins; D018995:Myosin Heavy Chains; D024501:Nonmuscle Myosin Type IIB; D011994:Recombinant Proteins; D000099163:Myosin Binding Protein C; C103815:nonmuscle myosin type IIB heavy chain