Creatine kinase: a role for arginine-95 in creatine binding and active site organization

Edmiston, Paul L.; Schavolt, Kristy L.; Kersteen, Elizabeth A.; Moore, Nichole R.; Borders, C.L.

doi:10.1016/s0167-4838(01)00159-5

Cited by 50 publications

(53 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Clearly, Y89Q has more specificity for D-arginine. These results are consistent with the idea that the 89th amino acid is the key residue for guanidino substrate recognition [20,21].…”

Section: Amino Acid 89supporting

confidence: 92%

“…The predicted structure indicated that the L-or D-arginine substrate is close to L64 in the N-terminal flexible loop (GS region) and N320 on the C-terminal flexible loop. Y89, which is proposed to be the key residue for guanidino substrate recognition [20,21], is also near the substrate arginines.…”

Section: Resultsmentioning

confidence: 99%

“…Amino acid residue-89 is proposed to be one of the key residues for recognition of guanidino substrates in phosphagen kinases [20,21]. Multiple alignments of amino acid sequences clearly indicated that the residue is strictly conserved as a specific amino acid in each phosphagen kinase: R in CK, Y in typical AK, I in GK, H in TK, and K in LK [24] (see Fig.…”

Section: Amino Acid 89mentioning

confidence: 97%

“…While this residue does not appear to be directly involved in substrate binding in the TSAC structures of CK and AK, it is located close to the guanidine substrate-binding site [14,15]. Site-directed mutagenesis studies of this residue using rabbit CK and Einenia LK have shown that it has a significant effect on enzymatic activity and guanidino substrate specificity [20,21]. A recent report investigating Danio CK and Nautilus AK also showed that the 89th residue plays a key role in organizing the hydrogen-bond network between the four dynamic domains [25] of AK and CK, thereby helping provide an appropriate active center for high catalytic turnover [26].…”

Section: Amino Acid 89mentioning

confidence: 99%

See 3 more Smart Citations

Identification of the key amino acid residues in Sabellastarte arginine kinase for distinguishing chiral guanidino substrates (d- and l-arginine)

Uda

Matumoto

Suzuki

2010

Journal of Molecular Catalysis B: Enzymatic

View full text Add to dashboard Cite

Arginine kinase (AK, EC 2.7.3.3) is a phosphagen kinase widely distributed in lower and higher invertebrates. Sabellastarte indica AKs (AK1 and AK2) are the only AKs that use D-arginine in addition to L-arginine as a substrate. Sabellastarte indica AKs are therefore clearly unrelated to typical AK. The purpose of this work was to identify the amino acid residues responsible for distinguishing the chirality of D-and L-arginine. The three-dimensional structure of Sabellastarte AK2 was predicted using the SWISS-MODEL automated modeling server. Two amino acids, L64 in the N-terminal flexible loop and N320 in the C-terminal flexible loop, were found to be closest to the substrate arginine. We introduced several mutations around the predicted binding site of the substrate arginine. In the L64I mutant, the affinity for L-arginine was greatly increased (9.5-fold that of the wild-type), whereas its affinity for D-arginine was increased 2.9-fold, indicating that the L64I mutant enzyme was more specific for L-arginine. On the other hand, the L64V mutant showed a 1.7-fold decrease in affinity for L-arginine, but unchanged affinity for D-arginine. Thus the L64V mutant enzyme was more specific for D-arginine. Since the replacement of amino acid residue L64 by I or V significantly affected the affinity for L-arginine or D-arginine, it can be concluded that amino acid 64 is a key residue for distinguishing D-and L-arginine. Seven mutants at position 320 (N320H, Q, D, E, R, K and A) had considerably reduced enzymatic activity (0.05 to 52% of the wild-type). The reduction in enzyme activity was more significant when D-arginine was the substrate rather than L-arginine, except for N320D, suggesting that the carbonyl oxygen of the Asn320 side chain forms a hydrogen bond with the α-amino nitrogen attached to the asymmetric carbon of the D-arginine substrate. In addition, we found that even a residue remote from the guanidino substrate binding site, such as G54 and Y89 in Sabellastarte 3 AK2, significantly affected guanidino substrate specificity.4

show abstract

“…Clearly, Y89Q has more specificity for D-arginine. These results are consistent with the idea that the 89th amino acid is the key residue for guanidino substrate recognition [20,21].…”

Section: Amino Acid 89supporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Amino Acid 89mentioning

confidence: 97%

Section: Amino Acid 89mentioning

confidence: 99%

See 2 more Smart Citations

Identification of the key amino acid residues in Sabellastarte arginine kinase for distinguishing chiral guanidino substrates (d- and l-arginine)

Uda

Matumoto

Suzuki

2010

Journal of Molecular Catalysis B: Enzymatic

View full text Add to dashboard Cite

show abstract

“…Edmiston et al [28] reported that R95Y, R95A and R95K mutants of rabbit muscle CK produced remarkably reduced CK activity. On the other hand, Uda and Suzuki [27] demonstrated a 70% decrease in V max for the Y95R mutant of Stichopus AK.…”

Section: Amino Acid 95 Is a Key Residue For Distinguishing Guanidinomentioning

confidence: 99%

Role of amino‐acid residue 95 in substrate specificity of phosphagen kinases

Tanaka

Suzuki

2004

FEBS Letters

View full text Add to dashboard Cite

The purpose of this study is to elucidate the mechanisms of guanidine substrate specificity in phosphagen kinases, including creatine kinase (CK), glycocyamine kinase (GK), lombricine kinase (LK), taurocyamine kinase (TK) and arginine kinase (AK). Among these enzymes, LK is unique in that it shows considerable enzyme activity for taurocyamine in addition to its original target substrate, lombricine. We earlier proposed several candidate amino acids associated with guanidine substrate recognition. Here, we focus on amino-acid residue 95, which is strictly conserved in phosphagen kinases: Arg in CK, Ile in GK, Lys in LK and Tyr in AK. This residue is not directly associated with substrate binding in CK and AK crystal structures, but it is located close to the binding site of the guanidine substrate. We replaced amino acid 95 Lys in LK isolated from earthworm Eisenia foetida with two amino acids, Arg or Tyr, expressed the modified enzymes in Escherichia coli as a fusion protein with maltose-binding protein, and determined the kinetic parameters. The K95R mutant enzyme showed a stronger affinity for both lombricine (K m = 0:74 mM and k cat =K m = 19:34 s À1 mM À1 ) and taurocyamine (K m = 2:67 and k cat =K m = 2:81), compared with those of the wild-type enzyme (K m = 5:33 and k cat =K m = 3:37 for lombricine, and K m = 15:31 and k cat =K m = 0:48 for taurocyamine). Enzyme activity of the other mutant, K95Y, was dramatically altered. The affinity for taurocyamine (K m = 1:93 and k cat =K m = 6:41) was enhanced remarkably and that for lombricine (K m = 14:2 and k cat =K m = 0:72) was largely decreased, indicating that this mutant functions as a taurocyamine kinase. This mutant also had a lower but significant enzyme activity for the substrate arginine (K m = 33:28 and k cat =K m = 0:01). These results suggest that Eisenia LK is an inherently flexible enzyme and that substrate specificity is strongly controlled by the aminoacid residue at position 95.

show abstract

Cold-adapted Features of Arginine Kinase from the Deep-sea Clam Calyptogena kaikoi

et al. 2011

View full text Add to dashboard Cite

The heterodont clam Calyptogena kaikoi, which inhabits depths exceeding 3,500 m where low ambient temperatures prevail, has an unusual two-domain arginine kinase (AK) with molecular mass of 80 kDa, twice that of typical AKs. The purpose of this work is to investigate the nature of the adaptations of this AK for functioning at low temperatures. Recombinant C. kaikoi AK constructs were expressed, and their two-substrate kinetic constants (k(cat), K(a), and K(ia)) were determined at 10°C and 25°C, respectively. When measured at 25°C, the K(ia) values were tenfold larger than those for corresponding K(a) values, while at 10°C, the K(ia) values decreased remarkably, but the K (a) values were almost unchanged. The Calyptogena two-domain enzyme has threefold higher catalytic efficiency, calculated by k (cat)/(K(a)(ARG)·K(ia)(ATP) ), at 10°C, than that at 25°C, reflecting adaptation for function at reduced ambient temperatures. The activation energy (E(a)) and thermodynamic parameters were determined for Calyptogena two-domain enzyme and compared with those of two-domain enzymes from mesophilic Corbicula and Anthopleura. The value for E(a) of Calyptogena enzyme were about half of those for mesophilic enzymes, and a larger decrease in entropy was observed in Calyptogena AK reaction. Although large decrease in entropy increases the ΔG(o‡) value and consequently lowers the k(cat) value, this is compensated with its lower E(a) value thereby minimizing the reduction in its k(cat) value. These thermodynamic properties, together with the kinetic ones, are also present in the separated domain 2 of the Calyptogena two-domain enzyme.

show abstract

Creatine kinase: a role for arginine-95 in creatine binding and active site organization

Cited by 50 publications

References 27 publications

Identification of the key amino acid residues in Sabellastarte arginine kinase for distinguishing chiral guanidino substrates (d- and l-arginine)

Identification of the key amino acid residues in Sabellastarte arginine kinase for distinguishing chiral guanidino substrates (d- and l-arginine)

Role of amino‐acid residue 95 in substrate specificity of phosphagen kinases

Cold-adapted Features of Arginine Kinase from the Deep-sea Clam Calyptogena kaikoi

Contact Info

Product

Resources

About