Charge changes in loop 2 affect the thermal unfolding of the myosin motor domain bound to F-actin.


Abstract

The thermal unfolding of Dictyostelium discoideum myosin head fragments with alterations in the actin-binding surface loop 2 was studied by differential scanning calorimetry. Lengthening of loop 2 without concomitant charge changes led to decreases in the transition temperature of not more than 1.8 degrees C. Insertions with multiple positive or negative charges had a stronger destabilizing effect and led to reductions in the thermal transition temperature of up to 3.7 degrees C. In the presence of nucleotide, most mutants displayed similar or higher transition temperatures than M765. Only constructs M765(11/+6) and M765(20/+12) with long positively charged inserts showed transition temperatures that were more than 2 degrees C below the values measured for M765 in the presence of ADP, ADP-V(i), and ADP-BeF(3). Interaction with F-actin in the presence of ADP shifted the thermal transition of M765 by 6 degrees C, from 49.1 to 55.1 degrees C. The actin-induced increase in thermal stability varied between 1.2 and 9.1 degrees C and showed a strong correlation with the mutant constructs' affinity for actin. Our results show that length and charge changes in loop 2 do not significantly affect nucleotide-induced structural changes in the myosin motor domain, but they affect structural changes that occur when the motor domain is strongly bound to actin and affect the coupling between the actin- and nucleotide-binding sites. Study holds ProTherm entries: 7855, 7856, 7857, 7858, 7859, 7860, 7861, 7862, 7863, 7864, 7865, 7866, 7867, 7868, 7869, 7870, 7871, 7872, 7873, 7874, 7875, 7876, 7877, 7878, 7879, 7880, 7881, 7882, 7883, 7884, 7885, 7886, 7887, 7888 Extra Details: actin-binding surface loop; insertions; nucleotide-induced,structural changes; nucleotide-binding sites

Submission Details

ID: rsLr3U9G

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:35 p.m.

Version: 1

Publication Details
Ponomarev MA;Furch M;Levitsky DI;Manstein DJ,Biochemistry (2000) Charge changes in loop 2 affect the thermal unfolding of the myosin motor domain bound to F-actin. PMID:10758002
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1W9I 2006-03-16 1.75 Myosin II Dictyostelium discoideum motor domain S456Y bound with MgADP-BeFx
3MYK 2010-05-26 1.84 Insights into the Importance of Hydrogen Bonding in the Gamma-Phosphate Binding Pocket of Myosin: Structural and Functional Studies of Ser236
1VOM 1996-12-23 1.9 COMPLEX BETWEEN DICTYOSTELIUM MYOSIN AND MGADP AND VANADATE AT 1.9A RESOLUTION
2AKA 2005-08-23 1.9 Structure of the nucleotide-free myosin II motor domain from Dictyostelium discoideum fused to the GTPase domain of dynamin 1 from Rattus norvegicus
1LVK 1998-01-28 1.9 X-RAY CRYSTAL STRUCTURE OF THE MG (DOT) 2'(3')-O-(N-METHYLANTHRANILOYL) NUCLEOTIDE BOUND TO DICTYOSTELIUM DISCOIDEUM MYOSIN MOTOR DOMAIN
1W9L 2006-03-16 1.95 Myosin II Dictyostelium discoideum motor domain S456E bound with MgADP-AlF4
1D0Z 2000-01-12 2.0 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH P-NITROPHENYL AMINOETHYLDIPHOSPHATE BERYLLIUM TRIFLUORIDE.
1YV3 2005-03-08 2.0 The structural basis of blebbistatin inhibition and specificity for myosin II
3MYL 2010-05-26 2.0 Insights into the Importance of Hydrogen Bonding in the Gamma-Phosphate Binding Pocket of Myosin: Structural and Functional Studies of Ser236
1D0Y 2000-01-12 2.0 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH O-NITROPHENYL AMINOETHYLDIPHOSPHATE BERYLLIUM FLUORIDE.
1D1A 2000-01-12 2.0 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH O,P-DINITROPHENYL AMINOETHYLDIPHOSPHATE BERYLLIUM TRIFLUORIDE.
1MMD 1996-08-17 2.0 TRUNCATED HEAD OF MYOSIN FROM DICTYOSTELIUM DISCOIDEUM COMPLEXED WITH MGADP-BEF3
1D1B 2000-01-12 2.0 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH O,P-DINITROPHENYL AMINOPROPYLDIPHOSPHATE BERYLLIUM TRIFLUORIDE.
3BZ7 2008-02-19 2.0 Crystal Structures of (S)-(-)-Blebbistatin Analogs bound to Dictyostelium discoideum myosin II
1W9J 2006-03-16 2.0 Myosin II Dictyostelium discoideum motor domain S456Y bound with MgADP-AlF4
1D0X 2000-01-12 2.0 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH M-NITROPHENYL AMINOETHYLDIPHOSPHATE BERYLLIUM TRIFLUORIDE.
3MYH 2010-05-26 2.01 Insights into the Importance of Hydrogen Bonding in the Gamma-Phosphate Binding Pocket of Myosin: Structural and Functional Studies of Ser236
1W9K 2006-03-16 2.05 Dictyostelium discoideum Myosin II motor domain S456E with bound MgADP-BeFx
1FMV 2000-11-22 2.1 CRYSTAL STRUCTURE OF THE APO MOTOR DOMAIN OF DICTYOSTELLIUM MYOSIN II
1MMG 1997-12-03 2.1 X-RAY STRUCTURES OF THE MGADP, MGATPGAMMAS, AND MGAMPPNP COMPLEXES OF THE DICTYOSTELIUM DISCOIDEUM MYOSIN MOTOR DOMAIN
1MMN 1997-12-03 2.1 X-RAY STRUCTURES OF THE MGADP, MGATPGAMMAS, AND MGAMPPNP COMPLEXES OF THE DICTYOSTELIUM DISCOIDEUM MYOSIN MOTOR DOMAIN
3BZ9 2008-02-19 2.1 Crystal Structures of (S)-(-)-Blebbistatin Analogs bound to Dictyostelium discoideum myosin II
1MMA 1997-12-03 2.1 X-RAY STRUCTURES OF THE MGADP, MGATPGAMMAS, AND MGAMPPNP COMPLEXES OF THE DICTYOSTELIUM DISCOIDEUM MYOSIN MOTOR DOMAIN
1FMW 2000-11-22 2.15 CRYSTAL STRUCTURE OF THE MGATP COMPLEX FOR THE MOTOR DOMAIN OF DICTYOSTELIUM MYOSIN II
4PJK 2015-04-29 2.15 Dicty myosin II R238E.E459R mutant (with ADP.Pi) in the Pi release state
3MJX 2011-04-27 2.2 Crystal structure of myosin-2 motor domain in complex with ADP-Metavanadate and blebbistatin
3BZ8 2008-02-19 2.2 Crystal Structures of (S)-(-)-Blebbistatin Analogs bound to Dictyostelium discoideum myosin II
3MNQ 2011-04-27 2.2 Crystal structure of myosin-2 motor domain in complex with ADP-metavanadate and resveratrol
2JJ9 2009-01-13 2.3 Crystal structure of myosin-2 in complex with ADP-metavanadate
1JX2 2001-11-07 2.3 CRYSTAL STRUCTURE OF THE NUCLEOTIDE-FREE DYNAMIN A GTPASE DOMAIN, DETERMINED AS MYOSIN FUSION
1D1C 2000-01-12 2.3 DICTYOSTELIUM MYOSIN S1DC (MOTOR DOMAIN FRAGMENT) COMPLEXED WITH N-METHYL-O-NITROPHENYL AMINOETHYLDIPHOSPHATE BERYLLIUM TRIFLUORIDE.
1JWY 2001-11-07 2.3 CRYSTAL STRUCTURE OF THE DYNAMIN A GTPASE DOMAIN COMPLEXED WITH GDP, DETERMINED AS MYOSIN FUSION
3MKD 2011-04-06 2.4 Crystal structure of myosin-2 dictyostelium discoideum motor domain S456Y mutant in complex with adp-orthovanadate
2XO8 2011-05-25 2.4 Crystal Structure of Myosin-2 in Complex with Tribromodichloropseudilin
2XEL 2011-06-15 2.5 Molecular Mechanism of Pentachloropseudilin Mediated Inhibition of Myosin Motor Activity
4AE3 2013-01-16 2.5 Crystal structure of ammosamide 272:myosin-2 motor domain complex
1MND 1996-08-17 2.6 TRUNCATED HEAD OF MYOSIN FROM DICTYOSTELIUM DISCOIDEUM COMPLEXED WITH MGADP-ALF4
2X9H 2011-05-11 2.7 CRYSTAL STRUCTURE OF MYOSIN-2 MOTOR DOMAIN IN COMPLEX WITH ADP- METAVANADATE AND PENTACHLOROCARBAZOLE
1MNE 1996-08-17 2.7 TRUNCATED HEAD OF MYOSIN FROM DICTYOSTELIUM DISCOIDEUM COMPLEXED WITH MG-PYROPHOSPHATE
2JHR 2009-01-13 2.8 Crystal structure of myosin-2 motor domain in complex with ADP- metavanadate and pentabromopseudilin
1G8X 2001-01-17 2.8 STRUCTURE OF A GENETICALLY ENGINEERED MOLECULAR MOTOR
2Y0R 2011-07-20 2.85 Structural basis for the allosteric interference of myosin function by mutants G680A and G680V of Dictyostelium myosin-2
2Y8I 2011-07-20 3.13 Structural basis for the allosteric interference of myosin function by mutants G680A and G680V of Dictyostelium myosin-2
2Y9E 2011-07-20 3.4 Structural basis for the allosteric interference of myosin function by mutants G680A and G680V of Dictyostelium myosin-2

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Myosin-2 heavy chain P08799 MYS2_DICDI