Unspecific Arginine Kinase of Molecular Weight 150 000. Purification and Properties

Abstract

Arginine kinase has been purified from body-wall muscle of a marine polychaetous annelid, Sabella pavonina. The preparation is homogeneous to analytical ultracentrifugation and disc electrophoresis on polyaorylamide gel. The enzyme is labile and undergoes a rapid decrease of its specific activity, even in the presence of protecting agents. The molecular weight of the native enzyme is 150000 f 5000, the sedimentation coefficient si,,, being found to be equal to 7.5.Unlike the enzyme from lobster muscle (mol. wt -43000) and that from Sipunculus nudus muscle (mol. w t -83000), the phosphokinase from Sabella pavonina muscle is not strictly specific towards L-arginine. The enzyme shows no optical isomer specificity and is slightly active on a number of analogues of L-arginine, the integrity of the guanidine and carboxylic groups of which is indispensable to their activity. The enzyme is strictly specific for the nucleotidic substrate ATP.Partial isotopic exchange and kinetic studies suggest that the reaction mechanism is somewhat different from that described for sea-water crustacean muscle arpinine kinase and more similar to that reported for other phosphagen phosphokinases.

The use of arginine analogues for investigating the functional organization of the arginine-binding site in lobster muscle arginine kinase. Role of the ‘essential’ thiol group

“…D-Arginine is not a substrate for the lobster enzyme (Virden & Watts, 1964). On the other hand it is of mechanistic interest that arginine kinases of flatworms (Turbellaria) and some polychaete annelids will phosphorylate D-arginine, the latter species at a rate similar to that found with the normal L-isomer (Virden & Watts, 1964;Robin et al, 1969Robin et al, , 1971Robin et al, , 1975. Among these groups, species with enzymes that phosphorylate arginine analogues modified at the a-amino group are also found (Virden & Watts, 1964).…”

Evolutionary variation between a monomer and a dimer arginine kinase. Purification of the enzyme from Holothuria forskali and a comparison of some properties with that from Homarus vulgaris

“…Thus co-operativity between the subsites binding substrates at one catalytic site appears to be an evolutionarily variable feature of arginine kinases not necessarily related to subunit interactions. Cheung (1973) observed subsite cooperativity with the monomeric honey-bee enzyme, and Moreland (1974) negative co-operativity between subsites with the monomeric enzyme from Pecten maximus, but it has not been found with the monomeric kinases of the king-crab, Limulus (Blethen, 1972), the Australian crayfish, Panulirus longipes (Smith & Morrison, 1969), or the tetrameric enzyme from the annelid worm Sabella pavonina (Robin et al, 1971). Both the honey bee, Pecten and Holothuria are all advanced forms, indicating that this form of co-operativity is a late feature ofmolecular evolution; indeed, it appears to be the rule with the mammalian creatine kinases (Watts, 1973), but these species are sufficiently well separated on the evolutionary tree to suggest that co-operativity, like subunit dimerization (Watts, 1971), has occurred more than once in the evolution of the phosphagen kinases.…”

The mechanism and modes of inhibition of arginine kinase from the cockroach (Periplaneta americana)

“…Buttlaire and Cohn (1974b) determined a K i of about 45 mM as a competitive inhibitor of AK from the American lobster. D-Arginine is neither a substrate nor an inhibitor of AK from crayfish, Panulirus longipes (Smith and Morrison, 1971) or Trypanosoma cruzi (Pereira et al, 2003); however, both L-arginine and D-arginine are equally effective substrates for an unusual 150 kDal AK found in the annelid Sabella pavonina (Robin et al, 1971). The present investigation shows that AK from the cockroach is significantly inhibited by D-arginine at a K i close to that of the K m for L-arginine by a mechanism of competitive inhibition.…”