Tryptophan fluorescence quenching in rabbit skeletal myosin rod

Chang, Yoke Chen; Ludescher, Richard D.

doi:10.1016/0301-4622(93)80041-g

Cited by 8 publications

(10 citation statements)

References 26 publications

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“…The Ksv values for monomers from a two-component dynamic analysis (eq 2) were 8.38 ± 0.24 and <0.04 M™1 with fractional amplitudes of 0.93 and 0.07, respectively (Table 1); analysis Using a twocomponent dynamic-static model (eq 3) gave identical results with a static quenching constant of zero. These results for monomer quenching obtained using filters were identical to previous data collected without filters (Chang & Ludescher, 1993), indicating that the use of filters did not compromise the experimental quenching results.…”

Section: Steadysupporting

confidence: 85%

“…b Stern-Volmer quenching constants from analysis of time-resolved quenching data from duplicate data sets. The data for rod monomers are from Chang and Ludescher (1993).c Calculated using = 3.0 ns (Szabo & Rayner, 1980). d Calculated using the long lifetime, = 5.40 ns (Chang & Ludescher, 1993).…”

Section: Steadymentioning

confidence: 99%

“…The data for rod monomers are from Chang and Ludescher (1993).c Calculated using = 3.0 ns (Szabo & Rayner, 1980). d Calculated using the long lifetime, = 5.40 ns (Chang & Ludescher, 1993). The errors were calculated by propagation of measured errors in Af,v and .…”

Section: Steadymentioning

confidence: 99%

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Local Conformation of Rabbit Skeletal Myosin Rod Filaments Probed by Intrinsic Tryptophan Fluorescence

Chang¹,

Ludescher²

1994

Biochemistry

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Rabbit skeletal myosin rod contains two tryptophan residues per chain (four per coiled-coil) that are located about 50 and 175 A from the N-terminus of the light meromyosin (LMM) region of rod. We have characterized the local polarity, excited-state photophysics, solvent accessibility, and rotational dynamics of these tryptophans in myosin rod filaments at 125 mM KCl using steady-state and time-resolved fluorescence techniques. The fluorescence decays were described using a complex bimodal distribution with a discrete long-lifetime component of 5.44 ns (amplitude of 0.51) and a Gaussian distribution of short lifetimes with mean of 0.105 ns and width of 2.15 ns (amplitude 0.49). The discrete long-lifetime species was efficiently quenched by the neutral quencher acrylamide with a bimolecular collision constant (kq) of 0.85 x 10(9) M-1 s-1. The emission spectrum, lifetime distribution, and quenching behavior of the tryptophans in myosin rod monomers (at 0.5 M KCl) were quite similar. Time-resolved anisotropy decays of the rod monomers and filaments exhibited nearly identical double-exponential decays to a constant. Each had a fast, subnanosecond component (amplitude 0.07), probably corresponding to fast wobble of the tryptophans on the coiled-coil surface, a slower, approximately 6-ns component (amplitude approximately 0.04), corresponding to an unidentified internal, segmental mode of motion of the coiled-coil, and a constant (r infinity) of 0.15.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Steadysupporting

confidence: 85%

Section: Steadymentioning

confidence: 99%

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Local Conformation of Rabbit Skeletal Myosin Rod Filaments Probed by Intrinsic Tryptophan Fluorescence

Chang¹,

Ludescher²

1994

Biochemistry

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“…The intrinsic fluorescence of tryptophan has been commonly employed for the detection of the tertiary structural changes of proteins because tryptophan fluorescence is sensitive to the surface hydrophobicity change of protein [ 27 , 28 , 29 ]. The reduced tryptophan fluorescence and red shifting of the peak serves as an indicator of protein unfolding, which leads to the exposure of tryptophan residues to a polar environment [ 17 ].…”

Section: Discussionmentioning

confidence: 99%

The Solubility and Structures of Porcine Myofibrillar Proteins under Low-Salt Processing Conditions as Affected by the Presence of L-Lysine

Wang

et al. 2022

Foods

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This study aimed to investigate the presence of L-lysine (Lys) on the solubility and structures of myofibrillar proteins (MFPs) at different ionic strengths. Porcine MFPs were incubated at 4 °C with various levels of ionic strengths (0.15, 0.3, or 0.6 M NaCl) with or without the presence of 20 or 40 mM Lys. After 24 h of incubation, MFP solubility and turbidity were determined, and the particle size distribution, circular dichroism spectra, and intrinsic tryptophan fluorescence of MFP were analyzed to obtain their secondary and tertiary structure. Results showed that the solubilization effects of Lys on MFPs are dependent on the ionic strength. Particularly, the presence of Lys could improve MFP solubility at 0.3 M, which resembles salt-reducing processing conditions. Concomitantly, the secondary and tertiary structures were observed to change as a result of the varying ionic strengths and the addition of Lys, including myofibril swelling, dissociation of myosin filaments, uncoiling of α-helix, and unfolding of the tertiary structure. The possible mechanisms underlying the solubilization effects of Lys on MFPs at low ionic strengths are discussed from the perspective of protein structural changes.

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“…There are two tryptophans in each native rod chain at positions 533 and 618 (near the middle) which are considered to be in different environments (Chang and Ludescher, 1993) and even in different protein domains (Lehrer and Maeda, 1994). The unfolding of myosin rod in Gdn/HCl was very fast and independent of the rod concentration.…”

Section: Discussionmentioning

confidence: 99%

Kinetics of Folding of the Myosin Rod

Béchet

Nozais

1997

European Journal of Biochemistry

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The kinetics of the unfolding and refolding of the myosin rod have been studied by fluorescence and circular dichroism techniques, at different concentrations of protein and guanidine hydrochloride. The unfolding of the myosin rod was fast and at least biphasic in 2-3 M denaturant, with an initial immediate phase followed by a slower low-amplitude first-order phase. The refolding of the rod in 0.4-2 M guanidine hydrochloride was also at least biphasic; an initial immediate phase preceded a slow second-order phase. At the final denaturant concentration of 0.8 M, the amplitude of the burst phase was weakly dependent on the protein concentration and the rate constant of the refolding slow phase was optimal. These data are incorporated into a folding mechanism with at least three states. The high rates of the first steps of unfolding and refolding may be relevant for the functioning of the native myosin molecule by allowing a transient separation of the two strands of the myosin tail.

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Tryptophan fluorescence quenching in rabbit skeletal myosin rod

Cited by 8 publications

References 26 publications

Local Conformation of Rabbit Skeletal Myosin Rod Filaments Probed by Intrinsic Tryptophan Fluorescence

Local Conformation of Rabbit Skeletal Myosin Rod Filaments Probed by Intrinsic Tryptophan Fluorescence

The Solubility and Structures of Porcine Myofibrillar Proteins under Low-Salt Processing Conditions as Affected by the Presence of L-Lysine

Kinetics of Folding of the Myosin Rod

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