Proximity and ligand-induced movement of interdomain residues in myosin subfragment 1 containing trapped MgADP and MgPPi probed by multifunctional cross-linking.

Rajasekharan, K. N.; Sivaramakrishnan, M; Burke, M. Desmond

doi:10.1016/s0021-9258(18)60945-7

Cited by 21 publications

(9 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using BPIA, we have shown that the cross-linking between SH-l and the 25-kDa region is nucleotide independent whereas that between SH-l and the 50-kDa region requires the presence of a Mg2+-nucleotide (Lu et al, 1986). Later similar results were obtained by Rajasekharan et al (1987) using the multifunctional cross-linker 4,4'-bis(7V-maleimido)benzophenone. It has also been reported that cross-links can form between SH-l and the 50-kDa region via 1,2,4-trinitrobenzene (Muno et al, 1987), and between SH-2 and the 25-kDa region via p-nitrophenyl iodoacetate in the absence of nucleotide (Hiratsuka, 1987).…”

Section: Discussionmentioning

confidence: 52%

Glutamic acid-88 is close to SH-1 in the tertiary structure of myosin subfragment-1

Lu¹,

Wong²

1989

Biochemistry

View full text Add to dashboard Cite

The thiol-specific photoactivatable reagent benzophenone iodoacetamide (BPIA) can be selectively incorporated into the most reactive thiol, SH-1, of myosin S1, and upon photolysis, an intramolecular cross-link is formed between SH-1 and the N-terminal 25-kDa region of S1. If a Mg2+-nucleotide is present during photolysis, cross-links can be formed either with the 25-kDa region or with the central 50-kDa region [Lu et al. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 6392]. Comparison of the peptide maps of cross-linked and un-cross-linked S1 heavy chains indicates that the segment located about 12-16 kDa from the N-terminus of the heavy chain can be cross-linked to SH-1 via BPIA independently of the presence of a nucleotide whereas the segment located 57-60 kDa from the N-terminus can be cross-linked to SH-1 only in the presence of a Mg2+-nucleotide [Sutoh & Lu (1987) Biochemistry 26, 4511]. In this report, S1 was labeled with radioactive BPIA, photolyzed in the absence of nucleotide, and then degraded with proteolytic enzymes. Peptides containing cross-links were isolated by liquid chromatography and subjected to amino acid sequence analyses. The results show that Glu-88 is the major site and Asp-89 and Met-92 are the minor sites involved in cross-linking with SH-1 (Cys-707) via BPIA. These residues are very near the reactive lysine residue (Lys-83) but relatively remote in the primary structure from the putative nucleotide binding region.

show abstract

Section: Discussionmentioning

confidence: 52%

Glutamic acid-88 is close to SH-1 in the tertiary structure of myosin subfragment-1

Lu¹,

Wong²

1989

Biochemistry

View full text Add to dashboard Cite

show abstract

“…There was a large difference in the extent of cross-linking when the photolysis was carried out in rigor as compared to when it was carried out in the presence of MgATP, suggesting that MgATP (and therefore most likely MgADP) binds to S-l in which both SHi and SH2 are modified. The following experiment similar to the one of Lu et al (1986) also suggests that MgADP binds to S-l in which both SH] and SH2 are modified (Rajasekharan et al, 1987): when S-l in which SH2 was modified with a photoreactive derivative of 4,4'-bis(A-maleimido)benzophenone (MBP) is exposed to light, the reagent cross-links SH2 to both 25and 50-kDa tryptic fragments of S-l. This is a strong suggestion that S-l modified with this reagent at SH2 binds MgADP.…”

Section: Discussionmentioning

confidence: 61%

Differential behavior of two cysteine residues on the myosin head in muscle fibers

Miyanishi¹,

Borejdo²

1989

Biochemistry

View full text Add to dashboard Cite

We have previously shown that the orientation of (iodoacetamido)tetramethylrhodamine labels on SH1 thiol of S-1 moieties changes when MgADP is added to the fibers in rigor [Borejdo, J., Assulin, O., Ando, T., & Putnam, S. (1982) J. Mol. Biol. 158, 391-414. Burghardt, T.P., Ando, T., & Borejdo, J. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 7515-7519]. Here we report the results of experiments in which the SH2 of S-1 was labeled with maleimidorhodamine. The specificity of modification of thiols was checked by measuring the stoichiometry of attached dye, by determining the extent of the decrease in EDTA (K+)- and Ca2+-ATPase activities, and by the localization of the dyes on peptides containing SH1 and/or SH2. Labeled S-1 was diffused into single glycerinated fibers of rabbit psoas muscle, and the orientation of chromophores was measured by fluorescence detected dichroism. The dye attached to SH1 was oriented at 65 degrees with respect to the fiber axis in rigor and at 51 degrees in the presence of MgADP, regardless of whether SH2 was modified or not. The dye on SH2 was oriented near 42 degrees both in the presence and in the absence of ADP, regardless of whether SH1 was modified or not. Our results show that rhodamine oriented differently when attached to SH2 compared with when attached to SH1 and that in the former placement it was not sensitive to MgADP. We think this indicates that the SH2-containing region has a mobility different from that of the SH1-containing region, i.e., that this is evidence for internal flexibility of S-1.

show abstract

“…In the absence of nucleotide, the site of the attachment was identified as Arg-239, proximal to the N-terminus (Agarwal et al, 1991). 4,4'-Dimaleimidobenzophenone, a Trifunctional Cross-Linker, In a related approach, 4,4'-dimaleimidobenzophenone (35, Chart 3) was employed as a photoactivatable, trifunctional, thiol-trapping reagent to monitor the conformational changes of SFi that take place during phosphate binding (Rajasekharan et al, 1987). A BP bridge was established between the thiol-1 and thiol-2 groups and photolytically labeled different regions, depending on whether or not MgADP (or MgPPi) was present in the binding site.…”

Section: Site-directed Photolabeling With Polypeptides Containing 4-b...mentioning

confidence: 99%

Benzophenone Photophores in Biochemistry

1994

View full text Add to dashboard Cite

The photoactivatable aryl ketone derivatives have been rediscovered as biochemical probes in the last 5 years. The expanding use of benzophenone (BP) photoprobes can be attributed to three distinct chemical and biochemical advantages. First, BPs are chemically more stable than diazo esters, aryl azides, and diazirines. Second, BPs can be manipulated in ambient light and can be activated at 350-360 nm, avoiding protein-damaging wavelengths. Third, BPs react preferentially with unreactive C-H bonds, even in the presence of solvent water and bulk nucleophiles. These three properties combine to produce highly efficient covalent modifications of macromolecules, frequently with remarkable site specificity. This Perspectives includes a brief review of BP photochemistry and a selection of specific applications of these photoprobes, which address questions in protein, nucleic acid, and lipid biochemistry.

show abstract

Proximity and ligand-induced movement of interdomain residues in myosin subfragment 1 containing trapped MgADP and MgPPi probed by multifunctional cross-linking.

Cited by 21 publications

References 48 publications

Glutamic acid-88 is close to SH-1 in the tertiary structure of myosin subfragment-1

Glutamic acid-88 is close to SH-1 in the tertiary structure of myosin subfragment-1

Differential behavior of two cysteine residues on the myosin head in muscle fibers

Benzophenone Photophores in Biochemistry

Contact Info

Product

Resources

About