Figure 5. The C-lobe of Ca 2 –CaM is... | Rockefeller University Press

Figure 5.

$Figure 5. The C-lobe of Ca2+–CaM is necessary and sufficient to interact with chicken TRPV1-ARD. (A–E) SEC elution profiles of chicken (Gallus gallus) TRPV1-ARD (GgV1ARD) mixed with various CaM mutants. A physical interaction was observed in the presence of 2 mM Ca2+ for wild-type CaM, CaM12, and CaM C-lobe, as indicated by a left-shift of the peak for the mixed proteins (60 µM GgV1ARD and 60 µM CaM; black) compared with elution volumes of the individual proteins (gray, GgV1ARD; dashed black, CaM). Shown are representative traces from two repeats. Wild-type CaM was used in A, followed by CaM mutated at all four Ca2+ sites (B, CaM1234), the two N-lobe Ca2+ sites (C, CaM12), or the two C-lobe Ca2+ sites (D, CaM34). (E) The isolated C-lobe of CaM (residues 76–148) interacts with chicken TRPV1-ARD by SEC. (F) Coomassie-stained gels from SEC runs in E. The lanes from left to right are fractions of increasing elution volume, from 10.5 to 15.5 ml.$

The C-lobe of Ca²⁺–CaM is necessary and sufficient to interact with chicken TRPV1-ARD. (A–E) SEC elution profiles of chicken (Gallus gallus) TRPV1-ARD (GgV1ARD) mixed with various CaM mutants. A physical interaction was observed in the presence of 2 mM Ca²⁺ for wild-type CaM, CaM12, and CaM C-lobe, as indicated by a left-shift of the peak for the mixed proteins (60 µM GgV1ARD and 60 µM CaM; black) compared with elution volumes of the individual proteins (gray, GgV1ARD; dashed black, CaM). Shown are representative traces from two repeats. Wild-type CaM was used in A, followed by CaM mutated at all four Ca²⁺ sites (B, CaM1234), the two N-lobe Ca²⁺ sites (C, CaM12), or the two C-lobe Ca²⁺ sites (D, CaM34). (E) The isolated C-lobe of CaM (residues 76–148) interacts with chicken TRPV1-ARD by SEC. (F) Coomassie-stained gels from SEC runs in E. The lanes from left to right are fractions of increasing elution volume, from 10.5 to 15.5 ml.

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