Origin of octameric creatine kinases

Abstract

Mitochondrial creatine kinase (MiCK) occurs primarily as an octameric form localized in the mitochondrial intermembrane compartment in vertebrate tissues and echinoderm spermatozoa (both deuterostome groups). The octameric quaternary structure is thought to play important functional and enzyme targeting roles. We have found that the spermatozoa of the protostome polychaete Chaetopterus variopedatus contain three distinct isoenzymes of creatine kinase (CK) termed CK1, CK2 and CK3. CK3 appears to be present only in the sperm head/midpiece complex where mitochondria are restricted and has a subunit relative molecular mass (M r ) of 43.4 kDa. Gel permeation chromatography using Superdex 200HR showed that CK3 has a native M r of 344.9 kDa indicating that this enzyme exists as an octamer. Electron micrographs of negatively stained CK3 preparations show structures which are virtually identical to those that have been seen for octameric vertebrate MiCK. The above observations show that CK3 from C. variopedatus displays great similarities to MiCKs from vertebrates and echinoderms. Octamerization of CK is not an advanced feature. The evolution of octameric subunit association is ancient and occurred prior to the divergence of protostomes and deuterostomes.z 1998 Federation of European Biochemical Societies.

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

“…8). Such a lack of functional coupling is well known for ancient mitochondrial arginine kinase (53), a close homologue of CK occurring in some protostomian arthropods and mollusks, such as the tobacco horn worm, Limulus polyphemus, or some crustaceans (4). Interestingly, the lower chordate Ciona shows a C-terminal MtCK sequence that is intermediate between Chaetopterus and vertebrates.…”

“…From a phylogenetic perspective, it is interesting to compare vertebrate MtCK with the MtCK of Ciona intestinalis, a marine chordate, 2 and the very ancient octameric MtCK of C. variopedatus, an invertebrate annelid worm (4,5,51). Chaetopterus MtCK, the witness of a very early divergence of octameric mitochondrial and dimeric cytosolic CK occurring more than 670 million years ago, has preserved a high sequence identity of about 71% with vertebrate MtCK (6).…”

“…Chaetopterus MtCK, the witness of a very early divergence of octameric mitochondrial and dimeric cytosolic CK occurring more than 670 million years ago, has preserved a high sequence identity of about 71% with vertebrate MtCK (6). Despite this similarity, some divergent properties have also been observed with Chaetopterus MtCK, as for example a higher stability of the octameric structure (4,51). It has also been speculated that this ancient MtCK displays very tight binding to CL due to its high pI of 9.7 and several basic residues in the C-terminal domain (5).…”

“…In most vertebrate tissues, cytosolic dimeric CK is coexpressed with a predominantly octameric mitochondrial isoenzyme (MtCK), sarcomeric MtCK (sMtCK) or ubiquitous uMtCK (2). This divergence is an early phylogenetic event since octameric MtCK is already present in the protostome marine worm Chaetopterus variopedatus (4,5) and has preserved its sequence through evolution (6). These facts already point to a specific functional role of the mitochondrial CK octamer.…”

Functional consequences of a gene duplication and fusion event in an arginine kinase

“…The resulting pellets were exhaustively dialyzed against diethylaminoethyl (DEAE) running buffer [10·mmol·l -1 Tris-HCl (pH 8.1), 0.5·mmol·l -1 EDTA, 1·mmol·l -1 dithiothreitol (DTT)] and then applied to a DEAE Sephacel column (Amersham Biotech, Piscataway, NJ, USA) and eluted with a salt gradient of 0-400·mmol·l -1 KCl. Peak DEAE fractions were used for native and subunit relative molecular mass (Mr) determinations by fast-preparative liquid chromatography (FPLC) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively, as described by Ellington et al (1998).…”