Fluorine-containing analogs of intermediates in the shikimate pathway

• Published in: Biochemistry (Easton) Vol. 15; no. 24; pp. 5315 - 5320
• Main Authors: Pilch, Paul F, Somerville, Ronald L
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.11.1976
• Subjects: 3-Deoxy-7-Phosphoheptulonate Synthase - metabolism; Aldehyde-Lyases - metabolism; Escherichia coli; Fluorine; Kinetics; Periodic Acid - pharmacology; Phosphoenolpyruvate - analogs & derivatives; Phosphoenolpyruvate - metabolism; Shikimic Acid - metabolism; Tetroses - metabolism
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00669a018

The phosphoenolpyruvate analogue (Z)-phosphoenol-3-fluoropyruvate is a substrate for phenylalanine-inhibitable 3-deoxy-D-arabino-heptulosonic acid-7-phosphate synthase from Escherichia coli. In the presence of excess erythrose 4-phosphate, apparent KM values of 65 and 38 muM were observed for phosphoenol-3-fluoropyruvate and phosphoenolpyruvate, respectively. Because the apparent Vmax for phosphoenol-3-fluoropyruvate is only 1.17% of that for phosphoenolpyruvate, one can study the former as an inhibitor of 3-deoxy-arabino-heptulosonic acid-7-phosphate synthase. Kinetic experiments showed phosphoenol-3-fluoropyruvate to be competitive with respect to phosphoenolpyruvate. Two distinguishable Ki values of 8 and 48 muM were obtained. The product (3S)-3-deoxy--3-fluoro-arabino-heptulosonic acid 7-phosphate was purified, characterized, and shown to act as a substrate for 5-dehydroquinate synthase. 3-Deoxy-3-fluoro-arabino-heptulosonic acid 7-phosphate, in contrast to 3-deoxy-arabino-heptulosonic acid 7-phosphate reacts slowly or not at all with reagents specific for 2-keto-3-deoxy sugars and is relatively resistant to oxidative cleavage by sodium periodate. The expected product of periodate oxidation, 3-fluoro-3-formylpyruvate, cannot be detected. This observation was clarified by studies with model compounds.