The Actin View of the Myosin Contact Sites

In blue: the putative contacts between actin and the myosin heavy chain; In green: the residues which probably make contacts with the light chains.

Table of Acto-Myosin Contacts



Residues 1 to 9 of actin sequence

Actin

Sequence

Comments Methods Refs
1-4 deletion reduces actin activated ATPase activity of myosin, no change in S1 binding site-directed mutants [1]
1-4 two extra charges increases actin-activated ATPase activity, no change in motility assay site-directed mutants [2]
1-4 Asps replaced by neutral residues: decrease in ATPase activity in driven motility assay site-directed mutants [3][4]
1-4 docking experiments between the Lorenz et al. F-actin model and Dictostelium S1 modelling of acto-S1 [5]
1-4 docking experiments between the Lorenz et al. F-actin model and chicken pectoralis S1 modelling of acto-S1 [6]
1-4 & 11 cross-linked to 20-50 KDa junction of S1 in the absence of MgATP (rigor) covalent cross-linking [7]
1-12 both EDC and pPDM actin monomer-S1-complexes can be incorporated into FA-S1 covalent cross-linking [8]
1-7 no effect of antibody on the binding of S1 in absence of MgATP antibodies [9]
1-7 antibody does not block weak binding of S1 in presence of MgATP at low salt.

at equi -molar ratios actin activated ATPase activity is 20%

 antibodies [10][11]

[12][13]
1-28 (probably 1-7) EDC cross-linked to EDANS probe produced 20-fold increase in MgATPase rate but little effect on rigor binding covalent cross-linking [14]
1-28 peptide competes with S1 for binding and activates S1 ATPase activity about the same as F-actin synthetic actin peptide [15]
1-28 glutaraldehyde and EEDQ link actin and S1 and elevates Mg-ATPase activity covalent cross-linking [14]
1-44 contact with S1 by carbodiimide in absence of MgATP

weak but specific effect on activationof S1 ATPase activity

 split actin, covalent cross-linking

synthetic peptide

 [16]

[17]
3-4 neutral and charge-shifting substitutions of beta-actin inhibit actomyosin ATPase activity,to a lesser extent inhibits the sliding motility site-directed mutants [18]

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Residues 10 to 39 of actin sequence

Actin

Sequence

Comments Methods Refs
1-12 both EDC and pPDM actin monomer-S1-complexes can be incorporated into FA-S1 covalent cross-linking [8]
1-28 (probably 1-7) EDC cross-linked to EDANS probe produced 20-fold increase in MgATPase rate but little effect on rigor binding covalent cross-linking [14]
1-28 peptide competes with S1 for binding and activates S1 ATPase activity about the same as F-actin synthetic actin peptide [15]
1-28 glutaraldehyde and EEDQ link actin and S1 and elevates Mg-ATPase activity covalent cross-linking [14]
1-44 contact with S1 by carbodiimide in absence of MgATP weak but specific effect on activation of S1 ATPase activity split actin covalent cross-linking

synthetic peptide

 [16]

[17]
10 S1 binds at or close to Cys-10 binding of S1 affects rotational motion of a synthetic peptide in the presence of MgATP 19F-NMR

synthetic peptide/NMR

 [19]

[20]

18-29 exert 80% inhibition of MgATPase, strong inhibition of S1 binding + MgATP but no effect on S1 binding in rigor state antibodies [21]
20-41 20-41 on actin directly bind to caldesmon and are also recognized by S1 1H-NMR [22]
24, 25 docking experiments between the Lorenz et al. F-actin model and S1 modelling acto-S1 [5][6]
24, 25 charge reversal mutant only moderately inhibits S1 binding, not motility site-directed mutants, motility assay [23]
28 this Arg can be crosslinked to S-1 covalent cross-linking [24]

 

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Residues 40 to 69 of actin sequence


Actin

Sequence

Comments

Methods

Refs
1-44 contact with S1 by carbodiimide in absence of MgATP

weak but specific effect on activation of S1 ATPase activity

 split actin covalent cross-linking

synthetic peptide

 [16]

[17]
20-41 20-41 on actin directly bind to caldesmon and are also recognized by S1 1H-NMR [22]
40-113 close to the myosin heavy chain covalent cross-linking [25]
40-113 EDC links S1 to the synthetic peptide 40-113. The complex still binds and releases FA but has decreased MgATPase activity covalent cross-linking [26]
45-52, 60-69 protected region when S1 bind digestion [27]
47-48 10-fold lower affinity for S1, 30% reduction in motility assay but no change in activation of S1 ATPase activity subtilisin cleavage [28][29]
48-67 glutaraldehyde cross-links Lys-50 to S1 covalent cross-linking [30]
48-113 contacts with residues 214-561 of S1 covalent cross-linking [31]
61 label is perturbed by S1 binding 19F-NMR [32]
61 pyridoxal 5'-phosphate labelled monomers do not affect the binding of S1 specific chemical modification [33]
61 intramonomer distance between Cys374 and Lys61 decreases when S1 binds FRET [34]
61-69 protective effect of S1(A2) on this region digestion by subtilisin [35]
69-nuc this distance in F-actin (22Å) increases (26Å) when S1 binds FRET [36]

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Residues 70 to 359 of actin sequence


Actin

Sequence

 Comments Methods Refs
40-113 close to the myosin heavy chain covalent cross-linking [25]
40-113 EDC links S1 to the synthetic peptide 40-113. The complex still binds and releases FA but has decreased MgATPase activity covalent cross-linking [26]
48-113 contacts with residues 214-561 of S1 covalent cross-linking [31]
79-92 make contacts with Dictyostelium S1 modelling of acto-S1 [5]
91-100 may contact S1 heavy chain modelling of acto-S1 [6]
95 this Arg can be crosslinked to S-1 covalent cross-linking [24]
96-103, 112-125 antibodies compete with S1 for binding to actin; peptides compete with S1; binding prevented by Ca-pyrophosphate. Two S1 loci on opposite sides of subdomain 1 antibodies, synthetic peptides [37]
99, 100 make contacts with Dictyostelium S1 modelling of acto-S1 [5]
99, 100 charge reversal disrupts motility assay site-directed mutants [23]
103 competition between S1 and an anti-actin antibody specific to an epitope including Thr103 antibodies [26]
105-120 filamin binds to 105-120 and is dissociated by S1 competitive binding [38]
144 makes contact with Dictyostelium S1 modelling of acto-S1 [5]
144-148 close to chicken S1 heavy chain modelling of acto-S1 [6]
332-333 make contacts with Dictyostelium S1 modelling of acto-S1 [5]
332-334 close contact with chicken S1 heavy chain modelling of acto-S1 [6]
338-348 antibodies to this sequence are rapidly eliminated by S1 binding ± MgATP synthetic peptide [39]
338-348 peptide displaces S1 in the absence of ATP (tight binding) but does not displace S1 when MgATP is present 1H-NMR, ATPase activity, synthetic peptide [40]
341,345, 349,352 make close contacts with either chicken or Dictyostelium S1 modelling of acto-S1 [5][6]
348-358 antibodies do not decrease A-M ATPase activity antibodies [39]

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Residues 360 to C-terminus of actin sequence

Actin

Sequence

 Comments Methods Refs
360,362, 363,374 C-terminal region of actin interacts directly with N-terminus of MLC1 1H-NMR [41]
360-372 C-terminus of actin is close to MLC1 tryptic peptide binding [37][38][42]
360-362, 363 binds to MLC1 covalent cross-linking [7]
360,361 not essential for force generation site-directed mutants [23]
363-364, 369-375 the C-terminus is indirectly involved in binding S1 digestion by chymotrypsin [43]
374 S1 binding quenches Cys-374-pyrene by 70% fluorescence quench [44]
374 is 45Å from Lys83 on the S1 heavy chain FRET [45]
374 is 49-53Å from SH1 of S1 heavy chain FRET [46]
374 is 51Å from SH1 of S1 heavy chain FRET [47]
374 is 60Å from SH1 of S1 heavy chain FRET [48]
374 is 51Å from ATPase site of S1 heavy chain FRET [49]
374 is 57Å from Cys177 of MLC1 FRET [50]
374 is 51Å from SH1 of S1 heavy chain FRET [47]
374 captured by EDC in acto-S1 complex covalent cross-linking [8]
374 spin label does not move when S1 rotates saturation transfer EPR [51]
Nuc is more than 62Å from SH1 of S1 heavy chain and 61-77Å from ATPase site of S1 FRET [46][49]
Nuc S1 binding induces conformational changes in the Nuc cleft of actin fluorescence quench [52]

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