Purification and properties of esterase-4 from Drosophila mojavensis

Molecular weight estimation of esterase isoenzymes in closely related Drosophila Species (Diptera: Drosophilidae) in non-denaturing polyacrylamide gel electrophoresis

“…Graphic analysis for both the enzymes demonstrated that they presented MW values between 81 and 91 kDa ( Table 2). Previous studies have estimated a MW between 85 and 95 kDa for EST-4 in D. mojavensis using gel filtration HPLC and Sephadex G-150 chromatrography (Pen et al, 1984). Our results also confirmed that EST-4 and EST-5 had very close hydrodynamic properties as observed by their relationships in native polyacrylamide gels (Figure 2 -A .…”

Gene Duplication and Subsequent Differentiation of Esterases in Cactophilic Drosophila Species

“…These values were different from those of D. melanogaster obtained by Healy et al (1991) (Table 4), which are very close to the MWs obtained by Pen et al (1984), which were between 85 and 95 kD for a variant of the EST-4 (with altered specificity to -naphthyl acetate) using gel filtration chromatography. Pen et al (1984) also used denaturing gel electrophoresis (SDS-PAGE) and obtained the MWs of the subunits of EST-4 as 62-64 kD. In another study, Pen et al (1986a) determined the MWs for the subunits of EST-5 as 64-66 kD.…”

“…The set of proteins known as esterases constitute one of the most heavily studied groups of isozymes. In the Drosophila mulleri complex, which is the subject of this study, esterases have been extensively studied in several species, including D. serido (Lapenta et al, 1995(Lapenta et al, , 1998, D. buzzatii (East, 1982;Barker, 1994;Lapenta et al, 1995Lapenta et al, , 1998, D. mojavensis (Zouros et al, 1982;Zouros & Van Delden, 1982;Pen et al, 1984Pen et al, , 1986aPen et al, , 1986bMateus et al, 2009), D. arizonae (Zouros et al, 1982;Ceron, 1988;Mateus et al, 2009), D. aldrichi (F. M. Johnson et al, 1968Kambysellis et al, 1968) and D. mulleri (F. M. Johnson et al, 1968;Kambysellis et al, 1968;Richardson & Smouse, 1976;Ceron, 1988). Zouros et al (1982) detected two esterases with different patterns of temporal and tissuespecific expression in Drosophila mojavensis and D. arizonae (formerly D. arizonensis).…”

“…Johnson, 1973Johnson, , 1974, and gene duplication has been used as one explanation for their evolution (Zouros et al, 1982;Pen et al, 1990;Mateus et al, 2009). Several studies on the changes in esterase activity during development in species of the D. repleta group have detected two main β-esterases that show different tissue-specific and temporal expression patterns (Zouros et al, 1982;East, 1982;Pen et al, 1984;Pen et al, 1986aPen et al, , 1986bPen et al, 1990;Mateus et al, 2009). One esterase, named EST-4, is present only in later third instar larvae and early pupae and has a high concentration in the carcass.…”

Evolutionary genetics ofDrosophila esterases

“…As with the a-preferring isozyme, the major/3-preferring esterase has been characterised biochemically from representatives of the virilis, repleta, melanogaster and obscura species groups (Narise & Hubby, 1966;Narise, 1973;Sasaki & Narise, 1978;Mane, Tepper & Richmond, 1983;East, 1984;Pen, Rongen & Beintema, 1984;Pen, Van Beeumen & Beintema, 1986;Morton & Singh, 1985;White, Mane & Richmond, 1988;Farmer & Carter, 1989). For each group this includes purification, at least partially, and in some cases to homogeneity (entailing about 200-300 fold purification, depending on the species).…”