Rotational dynamics of 4-aminobutyrate aminotransferase.

Abstract:

:The fluorescence dye 1-anilinonaphthalene-8-sulfonate (ANS) was used as a probe of non-polar binding sites in 4-aminobutyrate aminotransferase. ANS binds to a single binding site of the dimeric protein with a Kd of 6 microM. Nanosecond emission anisotropy measurements were performed on the ANS-enzyme in an effort to detect independent rotation of the subunits in the native enzyme. The observed rotational correlation time (phi = 65 ns) corresponds to the rotation of a rather rigid dimeric structure. The microenvironment surrounding the natural probe pyridoxal-5-P covalently bound to the dimeric structure was explored using 31P-NMR at 72.86 MHz. In the native enzyme, the pyridoxal-5-P 31P-chemical shift is pH-independent, indicating that the phosphate group is well protected from the solvent. The correlation time determined from the 31P-spectrum of the aminotransferase exceeds the value calculated for the hydrated spherical model (phi = 40 ns). It is concluded that the phosphate of the pyridoxal-5-P molecule is rigidly bound to the active site of 4-aminobutyrate aminotransferase.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Churchich JE,Kim DS,Schnackerz KD

doi

10.1016/0014-5793(83)80971-5

subject

Has Abstract

pub_date

1983-08-22 00:00:00

pages

221-5

issue

1-2

eissn

0014-5793

issn

1873-3468

pii

0014-5793(83)80971-5

journal_volume

160

pub_type

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