Chemical modification of the most reactive thiol group of rabbit skeletal muscle phosphofructokinase, to reduce its affinity toward substrate ATP and activating monovalent cations.

Abstract:

:The most reactive single thiol group of rabbit skeletal muscle phosphofructokinase per protomer was modified with the following thiol reagents: iodoacetamide, iodoacetate, 2-hydroxyethyl disulfide, 3,3'-dithiodipropionate, and glutathione disulfide. As a result of the modification, there was increase in not only the apparent activation constants of activating monovalent cations, NH4+ (about 3-, 9-, 12-, 20-, and 30-fold, respectively) and K+ (about 3-, 10-, 15-, 17-, and 20-fold, respectively), but also the apparent Km for ATP (about 3-, 10-, 15-, 100-, and 20-fold, respectively) without any significant change in maximum velocity or apparent Km for fructose 6-phosphate in the presence of high concentrations of NH4+. These results suggest that modification of the thiol group destabilizes the enzyme-monovalent cation-MgATP complex proposed by Suelter [Science (1970) 168, 789-795], causing an apparent loss in catalytic activity.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Nakajima Y,Nakamura K

subject

Has Abstract

pub_date

1991-02-01 00:00:00

pages

251-5

issue

2

eissn

0021-924X

issn

1756-2651

journal_volume

109

pub_type

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