Immunochemical and Protein-Chemical Studies of Class I Fructose 1, 6-Diphosphate Aldolases from Staphylococci

Abstract

The class I fructose 1,6-diphosphate aldolases of five staphylococcal strains were purified and characterized. All of these enzymes could be renatured to active structures after exposure to a temperature of 90°C. By using antisera against the enzymes, we measured the immunological cross-reactions by microcomplement fixation and activity inhibition. The amino acid compositions of the purified proteins were determined and compared. Our results provide further insight into the phylogenetical relationships among the staphylococcal species studied. Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus warneri, and Staphylococcus haemolyticus form a group of highly related species. Staphylococcus aureus was shown to be closely related to this group, whereas all of the other staphylococcal species showed distant relationships. Staphylococcus sciuri subsp. sciuri and S . sciuri subsp. lentus occupy a rather isolated position within the genus Staphylococcus and are related to each other at the species level.The fructose 1,6-diphosphate (FDP) aldolases which are usually found in bacteria and fungi are class I1 enzymes (31). These enzymes need a divalent metal ion for activity and are inhibited by chelating agents. In contrast, class I aldolases have been found in most staphylococci (9) and a few other bacteria (13, 20, 21, 35). Class I aldolases can be irreversibly inhibited by dihydroxyacetone phosphate in the presence of sodium borohydride (12). The class I FDP aldolases from staphylococci and peptococci appear to be similar in enzymatic kinetics and molecular weight. The presence of class I aldolases in staphylococci is a useful property for distinguishing these organisms from other members of the family Micrococcaceae (9). Both class I1 aldolase activity and class I aldolase activity have been found in Staphylococcus hyicus and Staphylococcus intermedius strains, but only a class I1 enzyme occurs in Staphylococcus caseolyticus (6). This difference provides a simple method for classifying these staphylococci within the genus. Furthermore, the phylogenetic relationships of staphylococci have been determined by deoxyribonucleic acid (DNA)-DNA (15, 17, 24, 29, 32) and DNA-ribonucleic acid (RNA) homology (14) studies, as well as by comparing 16s ribosomal RNAs (23) and immunological cross-reactions of catalases (30,32). In the present study we used the following two essentially different methods to reveal the phylot Present address:

Recommended minimal standards for description of new staphylococcal species

“…Another characteristic feature of most staphylococci is the occurrence of a class I FBP aldolase (Fischer et al, 1983;Gotz et al, 1979Gotz et al, , 1980. Most micrococci possess a class I1 FBP aldolase that contains an essential divalent cation whereas class I enzymes are found in animals and plants and form a Schiff-base intermediate.…”

Evaluation of a ribosomal RNA gene probe for the identification of species and subspecies within the genus Staphylococcus

“…; cell wall peptidoglycan and teichoic acid composition (End1 et al, 1983; Schumacher-Perdreau et al, 1983); bacteriolytic activity patterns (Varaldo & Satta, 1978); serotyping (Pillet & Orta, 1981); electrophoretic comparison of isofunctional enzymes (Gotz & Schleifer, 1976;Zimmerman & Kloos, 1976;Zimmerman, 1976); immunological relatedness of catalases (Schleifer et al, 1979) and fructose-l,6-diphosphate aldolases (Fischer et al, 1983); gel electrophoresis of cellular proteins (Clink & Pennington, 1987; Pennington et al, 1991) and penicillin-binding proteins (Fontana et subspecies. The isolates chosen were those which could be assigned to 7 Type strain.…”

Distribution of class I and class II fructose biphosphate aldolases in halophilic archaebacteria

“…Now, with the establishment of two classes of aldolase, immunological techniques by use of either the class I or class II aldolase and amino acid composition homologies can be further used to determine their relationships within the genus, as in the case of Staphylococcus[23]. Now, with the establishment of two classes of aldolase, immunological techniques by use of either the class I or class II aldolase and amino acid composition homologies can be further used to determine their relationships within the genus, as in the case of Staphylococcus[23].…”