Kinetic and thermodynamic properties of two barley thioredoxin h isozymes, HvTrxh1 and HvTrxh2.

Abstract:

:Barley thioredoxin h isozymes 1 (HvTrxh1) and barley thioredoxin h isozymes 2 (HvTrxh2) show distinct spatiotemporal distribution in germinating seeds. Using a novel approach involving measurement of bidirectional electron transfer rates between Escherichia coli thioredoxin, which exhibits redox-dependent fluorescence, and the barley isozymes, reaction kinetics and thermodynamic properties were readily determined. The reaction constants were approximately 60% higher for HvTrxh1 than HvTrxh2, while their redox potentials were very similar. The primary nucleophile, CysN, of the active site Trp-CysN-Gly-Pro-CysC motif has an apparent pKa of 7.6 in both isozymes, as found by iodoacetamide titration, but showed approximately 70% higher reactivity in HvTrxh1, suggesting significant functional difference between the isozymes.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Maeda K,Hägglund P,Björnberg O,Winther JR,Svensson B

doi

10.1016/j.febslet.2010.06.028

subject

Has Abstract

pub_date

2010-08-04 00:00:00

pages

3376-80

issue

15

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(10)00523-5

journal_volume

584

pub_type

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