Crystallization of Mitochondrial Creatine Kinase on Negatively Charged Lipid Layers

Schnyder, Thomas; Cyrklaff, Marek; Fuchs, Karl Hermann; Wallimann, Theo

doi:10.1006/jsbi.1994.1015

Cited by 29 publications

(17 citation statements)

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“…similar to that reported for CK crystals (McPherson, 1973;Schnyder et al, 1990Schnyder et al, , 1991.…”

Section: Purification Of a K From A Natural Source (Horseshoe Crabs) supporting

confidence: 89%

“…cyto-solic apo-CK (McPherson, 1973). and apo-Mi-CK (Schnyder et al ., 1990(Schnyder et al ., , 1991 have previously been obtained from other sources of enzyme, it was necessary to form the TSA complex for horseshoe crab AK. Formation of the complex is likely stabilizing a flexible protein (Lui & Cunningham, 1966;Reed & Cohn, 1972;Forstner et al, 1996) into a single conformation.…”

Section: Discussionmentioning

confidence: 99%

“…A year-long search for crystallization conditions was unsuccessful, testing general conditions (Jancarik & Kim, 1991) and those similar to ones successful for other phosphagen kinases (McPherson, 1973;Berthou et al, 1975;Burgess et al, 1978;Gilliland et al, 1983;Hershenson et al, 1986;Schnyder et al, 1990Schnyder et al, , 1991, with and without common additives (McPherson et al, 1986).…”

Section: Methodsmentioning

confidence: 99%

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Expression, purification from inclusion bodies, and crystal characterization of a transition state analog complex of arginine kinase: A model for studying phosphagen kinases

et al. 1997

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Phosphagen kinases catalyze the reversible transfer of a phosphoryl group between guanidino phosphate compounds and ADP, thereby regenerating ATP during bursts of cellular activity. Large quantities of highly pure arginine kinase (EC 2.7.3.3), the phosphagen kinase present in arthropods, have been isolated from E. coli, into which the cDNA for the horseshoe crab enzyme had been cloned. Purification involves size exclusion and anion exchange chromatographies applied in the denatured and refolded states. The recombinant enzyme has been crystallized as a transition state analog complex. Near complete native diffraction data have been collected to 1.86 A resolution. Substitution of a recombinant source for a natural one, improvement in the purification, and data collection at cry0 temperatures have all yielded significant improvements in diffraction. This reaction and those of the rest of the guanidino (or phosphagen) kinase family, including creatine kinase (CK), enable ATP to be quickly regenerated from storage phosphagens during bursts of cellular activity. In addition to this "temporal" buffering, it has been proposed that these enzymes also function in "spatial" buff-

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“…similar to that reported for CK crystals (McPherson, 1973;Schnyder et al, 1990Schnyder et al, , 1991.…”

Section: Purification Of a K From A Natural Source (Horseshoe Crabs) supporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Expression, purification from inclusion bodies, and crystal characterization of a transition state analog complex of arginine kinase: A model for studying phosphagen kinases

et al. 1997

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“…1A, "top" and "bottom" faces). Mainly, these faces were found attached to phospholipid membranes when analyzed by electron microscopy (47). The very C-terminal stretch of vertebrate MtCK contains three basic residues, the side chains of which protrude from the putative membrane binding face (Fig.…”

Section: Resultsmentioning

confidence: 99%

C-terminal Lysines Determine Phospholipid Interaction of Sarcomeric Mitochondrial Creatine Kinase

Schlattner

Gehring

Vernoux

et al. 2004

Journal of Biological Chemistry

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High affinity interaction between octameric mitochondrial creatine kinase (MtCK) and the phospholipid cardiolipin in the inner mitochondrial membrane plays an important role in metabolite channeling between MtCK and inner membrane adenylate translocator, which itself is tightly bound to cardiolipin. Three Cterminal basic residues revealed as putative cardiolipin anchors in the x-ray structures of MtCK and corresponding to lysines in human sarcomeric MtCK (sMtCK) were exchanged by in vitro mutagenesis (K369A/E, K379Q/A/E, K380Q/A/E) to yield double and triple mutants. sMtCK proteins were bacterially expressed, purified to homogeneity, and verified for structural integrity by enzymatic activity, gel filtration chromatography, and CD spectroscopy. Interaction with cardiolipin and other acidic phospholipids was quantitatively analyzed by light scattering, surface plasmon resonance, and fluorescence spectroscopy. All mutant sMtCKs showed a strong decrease in vesicle cross-linking, membrane affinity, binding capacity, membrane ordering capability, and binding-induced changes in protein structure as compared with wild type. These effects did not depend on the nature of the replacing amino acid but on the number of exchanged lysines. They were moderate for Lys-379/Lys-380 double mutants but pronounced for triple mutants, with a 30-fold lower membrane affinity and an entire lack of alterations in protein structure compared with wild-type sMtCK. However, even triple mutants partially maintained an increased order of cardiolipin-containing membranes. Thus, the three Cterminal lysines determine high affinity sMtCK/cardiolipin interaction and its effects on MtCK structure, whereas low level binding and some effect on membrane fluidity depend on other structural components. These results are discussed in regard to MtCK microcompartments and evolution.

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“…12,18 Scanning transmission electron microscopy and X-ray diffraction have provided a picture of the mtCK octamer. 9,[19][20][21][22] It is a highly ordered cube of about 9.3 Â 9.3 Â 8.6 nm, which is made up of four elongated banana-shaped dimers arranged along a 4-fold symmetry axis. The top and bottom faces perpendicular to this axis contain the basic residues-rich C-terminal ends, and are believed to be responsible for membrane binding via interactions with acidic phospholipids.…”

Section: Introductionmentioning

confidence: 99%

Interfacial behavior of cytoplasmic and mitochondrial creatine kinase oligomeric states

et al. 2005

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Adsorption to the air/water interface of isoenzymes of creatine kinase was investigated using surface pressure-area isotherms and Brewster angle microscopy (BAM) observations. Octameric mitochondrial creatine kinase (mtCK) exhibits a significant affinity for the air/water interface. Whatever the mode of formation of the interfacial film, i.e., injection of the protein in the subphase or spreading onto the buffer surface, the final arrangement and conformation adopted by mtCK molecules lead to a similar result. In contrast, the dimeric isoenzymes mtCK and cytosolic MMCK do not induce any surface pressure variation. However, when the subphase contains 0.3M NaCl, both isoenzymes adsorb to the interface. When treated with 0.8 or 3M GdnHCl, muscle creatine kinase (MMCK) becomes surface active and occupies a greater surface than mtCK. This result contrasts with previous observations, often derived from monomeric proteins, that their surface activity is increased upon unfolding. It underlines the possible influence exerted by the protein oligomeric state on its interfacial activity. At a subphase pH of 8.8, which corresponds to the pI of octameric mtCK, the profiles of the isotherms obtained with dimeric and octameric states and the resistance to compression of the protein monolayers are significantly affected when compared to those recorded at pH 7.4. These data suggest that the octamer is more hydrophobic than the dimer and may contribute to explaining why octamers bind to the inner mitochondrial membrane while dimers do not.

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Crystallization of Mitochondrial Creatine Kinase on Negatively Charged Lipid Layers

Cited by 29 publications

References 0 publications

Expression, purification from inclusion bodies, and crystal characterization of a transition state analog complex of arginine kinase: A model for studying phosphagen kinases

Expression, purification from inclusion bodies, and crystal characterization of a transition state analog complex of arginine kinase: A model for studying phosphagen kinases

C-terminal Lysines Determine Phospholipid Interaction of Sarcomeric Mitochondrial Creatine Kinase

Interfacial behavior of cytoplasmic and mitochondrial creatine kinase oligomeric states

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