Soybean root nodule acid phosphatase.
Acid phosphatases are ubiquitous enzymes that exhibit activity against a variety of substrates in vitro, although little is known about their intracellular function. In this study, we report the isolation, characterization, and partial sequence of the major acid phosphatase from soybean (Glycine max L.) root nodules. The phosphatase was purified predominantly as a heterodimer with subunits of 28 and 31 kD; homodimers of both subunits were also observed and exhibited phosphatase activity. In addition to the general phosphatase substrate, p-nitrophenyl phosphate, the heterodimeric form of the enzyme readily hydrolyzed 5'-nucleotides, flavin mononucleotide, and O-phospho-L-Tyr. Low or negligible activity was observed with ATP or polyphosphate. Purified nodule acid phosphatase was stimulated by magnesium, inhibited by calcium and EDTA, and competitively inhibited by cGMP and cAMP with apparent Ki values of 7 and 12 microM, respectively. Partial N-terminal and internal sequencing of the nodule acid phosphatase revealed homology to the soybean vegetative storage proteins. There was a 17-fold increase in enzyme activity and a noticeable increase in protein levels detected by immunoblotting methods during nodule development. Both of these parameters were low in young nodules and reached a peak in mature, functional nodules, suggesting that this enzyme is important for efficient nodule metabolism.
Penheiter
A R
AR
Department of Biochemistry, University of Nebraska, Lincoln 68588-0664, USA.
Duff
S M
SM
Sarath
G
G
D000135:Acid Phosphatase / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D018517:Plant Roots / Q000201:enzymology* / Q000254:growth & development / Q000382:microbiology; D012230:Rhizobiaceae; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000201:enzymology* / Q000254:growth & development / Q000382:microbiology; D013379:Substrate Specificity; D013559:Symbiosis
D002413:Cations, Divalent; D002614:Chelating Agents; D000135:Acid Phosphatase