8ENC

Helical reconstruction of the human cardiac actin-tropomyosin-myosin loop 4 7G mutant complex

Summary for 8ENC

• Entry DOI: 10.2210/pdb8enc/pdb
• EMDB information: 28270
• Descriptor: Myosin-7, Actin, alpha cardiac muscle 1, Tropomyosin alpha-1 chain, ... (5 entities in total)
• Functional Keywords: actin, tropomyosin, myosin, cardiac, motor protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 8
• Total formula weight: 501040.56

Authors

Doran, M.H.,Lehman, W.,Rynkiewicz, M.J. (deposition date: 2022-09-29, release date: 2022-11-23, Last modification date: 2024-06-19)

Primary citation

Doran, M.H.,Rynkiewicz, M.J.,Pavadai, E.,Bodt, S.M.L.,Rasicci, D.,Moore, J.R.,Yengo, C.M.,Bullitt, E.,Lehman, W.
Myosin loop-4 is critical for optimal tropomyosin repositioning on actin during muscle activation and relaxation.
J.Gen.Physiol., 155:-, 2023

PubMed Abstract: During force-generating steps of the muscle crossbridge cycle, the tip of the myosin motor, specifically loop-4, contacts the tropomyosin cable of actin filaments. In the current study, we determined the corresponding effect of myosin loop-4 on the regulatory positioning of tropomyosin on actin. To accomplish this, we compared high-resolution cryo-EM structures of myosin S1-decorated thin filaments containing either wild-type or a loop-4 mutant construct, where the seven-residue portion of myosin loop-4 that contacts tropomyosin was replaced by glycine residues, thus removing polar side chains from residues 366-372. Cryo-EM analysis of fully decorated actin-tropomyosin filaments with wild-type and mutant S1, yielded 3.4-3.6 Å resolution reconstructions, with even higher definition at the actin-myosin interface. Loop-4 densities both in wild-type and mutant S1 were clearly identified, and side chains were resolved in the wild-type structure. Aside from loop-4, actin and myosin structural domains were indistinguishable from each other when filaments were decorated with either mutant or wild-type S1. In marked contrast, the position of tropomyosin on actin in the two reconstructions differed by 3 to 4 Å. In maps of filaments containing the mutant, tropomyosin was located closer to the myosin-head and thus moved in the direction of the C-state conformation adopted by myosin-free thin filaments. Complementary interaction energy measurements showed that tropomyosin in the mutant thin filaments sits on actin in a local energy minimum, whereas tropomyosin is positioned by wild-type S1 in an energetically unfavorable location. We propose that the high potential energy associated with tropomyosin positioning in wild-type filaments favors an effective transition to B- and C-states following release of myosin from the thin filaments during relaxation.

PubMed: 36459134
DOI: 10.1085/jgp.202213274

Experimental method

ELECTRON MICROSCOPY (3.6 Å)