Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues

Laura Moschcovich, Y. Michael Peyser, Claudio Salomon, Thomas P Burghardt, Andras Muhlrad

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

2-[(2-Nitrophenyl)amino]ethyl triphosphate (NPhAETP) is the smallest ATP analogue that serves as a substrate for the actin-activated ATPase of myosin subfragment 1 (S1) and supports the development of active tension in skinned fibers. 2-(Phenylamino)ethyl triphosphate (PhAETP), in which the nitro group on the phenyl ring of NPhAETP is substituted by a H atom, is also a substrate of the actin-activated ATPase but does not support active tension [Wang, D., Pate, E., Cooke, R., and Yount, R. (1993) J. Muscle Res. Cell Motil. 14, 484- 497]. We compared the S1-catalyzed hydrolysis of these analogues, their ability to support the formation of stable complexes with S1 and phosphate analogues, and their effect on S1 conformation. The analogues were hydrolyzed by S1 under various conditions both in the presence and in the absence of actin. In some cases, the effects of the two analogues are similar to each other and to those of ATP; they protect S1 from heat denaturation at 40 °C and inhibit the formation of the N-terminal 29 kDa fragment during the tryptic digestion of S1 and the modification of Lys-83 with trinitrobenzene sulfonate. However, in other cases, the effect of the two analogues is different; the effect of NPhAETP resembles that of ATP. NPhAETP and ATP decrease while PhAETP increases the rate of reaction of the SH1 thiol (Cys- 707) with coumarin maleimide. The diphosphate forms of the two analogues induce a much smaller change in the near-UV CD spectrum of S1 than ADP. NPhAEDP forms stable complexes with S1 in the presence of beryllium fluoride (BeF(x)), aluminum fluoride (AlF4 -), or vanadate (Vi) phosphate analogues, while the S1·PhAEDP complex is stable in the presence BeF(x) but much less stable with AlF4 - and Vi. These results indicate that the S1·PhAEDP·P1 state is poorly populated during the PhAETP hydrolysis. The models of the atomic structure of S1 complexed by the two analogues show that PhAETP, unlike NPhAETP or ATP, does not form a H bond with Tyr-134 in S1, which is the probable structural reason of the lack of tension development, with PhAETP as the substrate.

Original languageEnglish (US)
Pages (from-to)15137-15143
Number of pages7
JournalBiochemistry
Volume37
Issue number43
DOIs
StatePublished - Oct 27 1998

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Myosin Subfragments
Adenosine Triphosphate
Myosins
Hydrolysis
Substrates
Phosphates
Trinitrobenzenesulfonic Acid
triphosphoric acid
Denaturation
Vanadates
Diphosphates
Sulfhydryl Compounds
Adenosine Diphosphate
Muscle Cells
Muscle
Conformations
Actins
Digestion
Hot Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Moschcovich, L., Michael Peyser, Y., Salomon, C., Burghardt, T. P., & Muhlrad, A. (1998). Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues. Biochemistry, 37(43), 15137-15143. https://doi.org/10.1021/bi980605w

Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues. / Moschcovich, Laura; Michael Peyser, Y.; Salomon, Claudio; Burghardt, Thomas P; Muhlrad, Andras.

In: Biochemistry, Vol. 37, No. 43, 27.10.1998, p. 15137-15143.

Research output: Contribution to journalArticle

Moschcovich, L, Michael Peyser, Y, Salomon, C, Burghardt, TP & Muhlrad, A 1998, 'Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues', Biochemistry, vol. 37, no. 43, pp. 15137-15143. https://doi.org/10.1021/bi980605w
Moschcovich L, Michael Peyser Y, Salomon C, Burghardt TP, Muhlrad A. Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues. Biochemistry. 1998 Oct 27;37(43):15137-15143. https://doi.org/10.1021/bi980605w
Moschcovich, Laura ; Michael Peyser, Y. ; Salomon, Claudio ; Burghardt, Thomas P ; Muhlrad, Andras. / Interaction of myosin subfragment 1 with two non-nucleotide ATP analogues. In: Biochemistry. 1998 ; Vol. 37, No. 43. pp. 15137-15143.
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N2 - 2-[(2-Nitrophenyl)amino]ethyl triphosphate (NPhAETP) is the smallest ATP analogue that serves as a substrate for the actin-activated ATPase of myosin subfragment 1 (S1) and supports the development of active tension in skinned fibers. 2-(Phenylamino)ethyl triphosphate (PhAETP), in which the nitro group on the phenyl ring of NPhAETP is substituted by a H atom, is also a substrate of the actin-activated ATPase but does not support active tension [Wang, D., Pate, E., Cooke, R., and Yount, R. (1993) J. Muscle Res. Cell Motil. 14, 484- 497]. We compared the S1-catalyzed hydrolysis of these analogues, their ability to support the formation of stable complexes with S1 and phosphate analogues, and their effect on S1 conformation. The analogues were hydrolyzed by S1 under various conditions both in the presence and in the absence of actin. In some cases, the effects of the two analogues are similar to each other and to those of ATP; they protect S1 from heat denaturation at 40 °C and inhibit the formation of the N-terminal 29 kDa fragment during the tryptic digestion of S1 and the modification of Lys-83 with trinitrobenzene sulfonate. However, in other cases, the effect of the two analogues is different; the effect of NPhAETP resembles that of ATP. NPhAETP and ATP decrease while PhAETP increases the rate of reaction of the SH1 thiol (Cys- 707) with coumarin maleimide. The diphosphate forms of the two analogues induce a much smaller change in the near-UV CD spectrum of S1 than ADP. NPhAEDP forms stable complexes with S1 in the presence of beryllium fluoride (BeF(x)), aluminum fluoride (AlF4 -), or vanadate (Vi) phosphate analogues, while the S1·PhAEDP complex is stable in the presence BeF(x) but much less stable with AlF4 - and Vi. These results indicate that the S1·PhAEDP·P1 state is poorly populated during the PhAETP hydrolysis. The models of the atomic structure of S1 complexed by the two analogues show that PhAETP, unlike NPhAETP or ATP, does not form a H bond with Tyr-134 in S1, which is the probable structural reason of the lack of tension development, with PhAETP as the substrate.

AB - 2-[(2-Nitrophenyl)amino]ethyl triphosphate (NPhAETP) is the smallest ATP analogue that serves as a substrate for the actin-activated ATPase of myosin subfragment 1 (S1) and supports the development of active tension in skinned fibers. 2-(Phenylamino)ethyl triphosphate (PhAETP), in which the nitro group on the phenyl ring of NPhAETP is substituted by a H atom, is also a substrate of the actin-activated ATPase but does not support active tension [Wang, D., Pate, E., Cooke, R., and Yount, R. (1993) J. Muscle Res. Cell Motil. 14, 484- 497]. We compared the S1-catalyzed hydrolysis of these analogues, their ability to support the formation of stable complexes with S1 and phosphate analogues, and their effect on S1 conformation. The analogues were hydrolyzed by S1 under various conditions both in the presence and in the absence of actin. In some cases, the effects of the two analogues are similar to each other and to those of ATP; they protect S1 from heat denaturation at 40 °C and inhibit the formation of the N-terminal 29 kDa fragment during the tryptic digestion of S1 and the modification of Lys-83 with trinitrobenzene sulfonate. However, in other cases, the effect of the two analogues is different; the effect of NPhAETP resembles that of ATP. NPhAETP and ATP decrease while PhAETP increases the rate of reaction of the SH1 thiol (Cys- 707) with coumarin maleimide. The diphosphate forms of the two analogues induce a much smaller change in the near-UV CD spectrum of S1 than ADP. NPhAEDP forms stable complexes with S1 in the presence of beryllium fluoride (BeF(x)), aluminum fluoride (AlF4 -), or vanadate (Vi) phosphate analogues, while the S1·PhAEDP complex is stable in the presence BeF(x) but much less stable with AlF4 - and Vi. These results indicate that the S1·PhAEDP·P1 state is poorly populated during the PhAETP hydrolysis. The models of the atomic structure of S1 complexed by the two analogues show that PhAETP, unlike NPhAETP or ATP, does not form a H bond with Tyr-134 in S1, which is the probable structural reason of the lack of tension development, with PhAETP as the substrate.

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