Location of two contact sites between human smooth muscle caldesmon and Ca(2+)-calmodulin.

Marston, Steven B.; Fraser, Iain D. C.; Huber, P.; Pritchard, Kevin; Gusev, Nikolai B.; Török, Katalin

doi:10.1016/s0021-9258(17)37170-3

Cited by 58 publications

(28 citation statements)

References 33 publications

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“…Synthetic Peptides-«Three synthetic peptides were used in this study: GS17C (from Gly-651 to Ser-667 (GVRNIKSMWEKGNVFSS-C», VG29C (from Val-685 to Gly-713 (VSSRINEWLTKTPEGNKSPAP-KPSDLRPG-C)), and MG56C (from Met-658 to Gly-713 (GVRNIKS-MWEKGNVFSSPGGTGTPNKETAGLKVGVSSRINEWLTKTPEGNK-SPAPKPSDLRPG-C)); each contains a Cys residue at the C terminus in addition to the CaD sequence. GS17C contains site A (singly underlined, Met-658 to Ser-666, see Zhan et al (1991», VG29C encompasses site B for CaM (doubly underlined, Ser-687 to Lys-695) on the basis of the reports by Marston et al (1994) and Mezgueldi et al (1994), and MG56C contains both site A and site B. All peptides were synthesized on an automated ABI peptide synthesizer (model 431A) using F (N-(9-fluorenyl)methoxycarbonyl) chemistry.…”

Section: Methodsmentioning

confidence: 99%

“…1), also binds CaM in a Ca 2 + -dependent manner (Mezgueldi et al, 1994) supports this view. Furthermore, Marston et al (1994) found that although a site A-containing peptide (M73, from Ser-657 to Gly-670) did bind CaM, it could not compete with CaD for CaM and did not restore the inhibition of actomyosin ATPase activity, whereas a site B-containing fragment H2 (from Thr-626 to Leu-TlO) did. From these results, they concluded that it is site B, not site A, that is functionally relevant for CaD's action (Marston et al, 1994).…”

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

“…One may argue that since site A is at the very end ofthe N terminus ofMG56C, it may not fold properly without the flanking sequence, thus lowering the affinity for CaM as compared with GS17C. This, however, seems unlikely, because M73, which begins at the same residue as MG56C, does not exhibit a lowered affinity for CaM (Marston et al, 1994) (see Fig. 1).…”

Section: Summary Of Binding Parameters For Cam•peptide Interactionsmentioning

confidence: 99%

“…More recently, Marston et al(1994) suggested that there is another CaM-binding site (referred to as "site B") adjacent to the GS17C sequence (site A) and that the two sites do not compete with each other for CaM. The fact that a synthetic peptide, NK21 (from Asn-675 to Lys-695, see Fig.…”

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

“…Amino acid sequence and numbering of residues are according to chicken gizzard CaD (Bryan et al, 1989). The peptides/fragments indicated are: GS17C, VG29C, and MG56C (this work); M73, H2, and H9 (Marston et al, 1994); and NK21 (Mezgueldi et al, 1994).…”

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

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Identification of the Functionally Relevant Calmodulin Binding Site in Smooth Muscle Caldesmon

Zhuang

Wang

1995

Journal of Biological Chemistry

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The C-terminal region of smooth muscle caldesmon (CaD) interacts with calmodulin (CaM) and reverses CaD's inhibitory effect on the actomyosin ATPase activity. We have previously shown that the major CaM-binding site (site A) in this region is within the segment from Met-658 to Ser-666 (Zhan, Q., Wong, S. S., and Wang, C.-L. A. (1991) J. Biol. Chem. 266, 21810-21814). Recently, another segment (site B), Asn-675 to Lys-695, was reported to bind CaM (Mezgueldi, M., Derancourt, J., Calas, B., Kassab, R., and Fattoum, A. (1994) J. Biol. Chem. 269, 12824-12832). To assess the functional relevance of these two putative CaM-binding sites, we have examined three synthetic peptides regarding their effects on CaM's ability to reverse CaD-induced inhibition of actomyosin ATPase activity: GS17C (Gly-651 to Ser-667), VG29C (Val-685 to Gly-713), each containing one CaM-binding site, and MG56C (Met-658 to Gly-713), which contains both sites. We found that although VG29C did bind CaM, its affinity was weakened by GS17C, and it failed to compete with CaD for CaM under the conditions where GS17C effectively displaced CaD from CaM. MG56C had an effect similar to that of GS17C. These experiments demonstrated that site A for CaM binding is involved in regulating the inhibitory property of CaD.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

Section: Summary Of Binding Parameters For Cam•peptide Interactionsmentioning

confidence: 99%

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

Section: Identification Of the Functionally Relevant Calmodulin Binding Site In Smooth Muscle Caldesmon*mentioning

confidence: 99%

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Identification of the Functionally Relevant Calmodulin Binding Site in Smooth Muscle Caldesmon

Zhuang

Wang

1995

Journal of Biological Chemistry

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Two domains of interaction with calcium binding proteins can be mapped using fragments of calponin

et al. 1994

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Native calponin is able to bind 2 mol of calcium binding protein (CaBP) per mole calponin. This study extends this observation to define the 2 domains of interaction, one of which is near the actin binding site, and the other in the amino-terminal region of calponin. Also, the first evidence for a differentiation in the response of calponin to interaction with caltropin versus calmodulin is demonstrated. The binding of caltropin to cleavage and recombinant fragments of calponin was determined by 3 techniques: tryptophan fluorescence of the fragments, CD measurements to determine secondary structure changes, and analytical ultracentrifugation. In order to delineate the sites of interaction, 3 fragments of calponin have been studied. From a cyanogen bromide cleavage of calponin, residues 2-51 were isolated. This fragment is shown to bind to CaBPs and the affinity for caltropin is slightly higher than that for calmodulin. A carboxyl-terminal truncated mutant of calponin comprising residues 1-228 (CP 1-228) has been produced by recombinant techniques. Analytical ultracentrifugation has shown that C P 1-228, like the parent calponin, is able to bind 2 mol of caltropin per mol of 1-228 in a Ca'+-dependent fashion, indicating that there is a second site of interaction between residues 52-228. Temperature denaturation of the carboxylterminal truncated fragment compared with whole calponin show that the carboxyl-terminal region does not change the temperature at which calponin melts; however, there is greater residual secondary structure with whole calponin versus the fragment. A second mutant produced through recombinant techniques comprises residues 45-228 and is also able to bind caltropin, thus mapping the location of the second site of interaction to near the actin binding site.

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Regulation of cross-bridge cycling by Ca2+ in smooth muscle

Arner

Pfitzer

1999

Reviews of Physiology Biochemistry and Pharmacology, Volume 134

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Location of two contact sites between human smooth muscle caldesmon and Ca(2+)-calmodulin.

Cited by 58 publications

References 33 publications

Identification of the Functionally Relevant Calmodulin Binding Site in Smooth Muscle Caldesmon

Identification of the Functionally Relevant Calmodulin Binding Site in Smooth Muscle Caldesmon

Two domains of interaction with calcium binding proteins can be mapped using fragments of calponin

Regulation of cross-bridge cycling by Ca2+ in smooth muscle

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