Plant Physiol.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Plant Physiology 84:654-657 (1987)
© 1987 American Society of Plant Biologists

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Web of Science (28)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Streit, L.
Right arrow Articles by Harper, J. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Streit, L.
Right arrow Articles by Harper, J. E.
Agricola
Right arrow Articles by Streit, L.
Right arrow Articles by Harper, J. E.
Metabolism and Enzymology

A Method for the Separation and Partial Purification of the Three Forms of Nitrate Reductase Present in Wild-Type Soybean Leaves 1

Luc Streit2, Barry A. Martin3 and James E. Harper

Department of Agronomy, University of Illinois, 1102 S. Goodwin Ave., Urbana, Illinois 61801, United States Department of Agriculture-Agricultural Research Service, University of Illinois, 1102 S. Goodwin Ave., Urbana, Illinois 61801

A rapid and simple purification method was used to separate and purify nitrate reductases (NR) from Williams soybean leaves. Blue Sepharose columns were sequentially eluted with 50 millimolar NADPH and 50 millimolar NADH, thus separating NAD(P)H:NR from NADH:NRs. Subsequent purification of the collected peaks on a fast protein liquid chromatography-Mono Q column enabled separation of two NADH:NRs. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the subunit relative molecular mass for all three NR forms (constitutive NAD(P)H:NR [pH 6.5], EC 1.6.6.2; constitutive NADH:NR [pH 6.5], EC not assigned; and inducible NADH:NR [pH 7.5], EC 1.6.6.1) was approximately 107 to 109 kilodaltons. All three NRs showed similar spectra with absorption maxima at 413 and 273 nanometers in the oxidized state, and with the characteristics of a cytochrome b type heme upon reduction with NADH (absorption maxima at 556, 527, and 424 nanometers). The technique developed provides an improved separation of the three NR forms from soybean leaves. The similarity of the NRs with regard to their cytochrome b556 type heme content and in relative molecular mass indicated that other differences must exist to account for the different kinetic and physical properties previously reported.

2 Present address: Ciba-Geigy Ltd., Postfach, 4002, Basle, Switzerland.

3 Present address: Pioneer HiBred International, Inc., Plant Breeding Division, P. O. Box 85, Johnston, IA 50131-0085.

1 Supported by Competitive Research Grant No. 82-CRCR-1-1038.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
ASPB Publications PLANT PHYSIOLOGY® THE PLANT CELL
Copyright © 1987 by the American Society of Plant Biologists