Alternative arrangements of catalytic residues at the active sites of restriction enzymes.

Abstract:

:A catalytic sequence motif PDX10-30(E/D)XK is found in many restriction enzymes. On the basis of sequence similarities and mapping of the conserved residues to the crystal structure of NgoMIV we suggest that residues D160, K182, R186, R188 and E195 contribute to the catalytic/DNA binding site of the Ecl18kI restriction endonuclease. Mutational analysis confirms the functional significance of the conserved residues of Ecl18kI. Therefore, we conclude that the active site motif 159VDX21KX12E of Ecl18kI differs from the canonical PDX10-30(E/D)XK motif characteristic for most of the restriction enzymes. Moreover, we propose that two subfamilies of endonucleases Ecl18kI/PspGI/EcoRII and Cfr10I/Bse634I/NgoMIV, specific, respectively, for CCNGG/CCWGG and RCCGGY/GCCGGC sites, share conserved active site architecture and DNA binding elements.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Tamulaitis G,Solonin AS,Siksnys V

doi

10.1016/s0014-5793(02)02621-2

keywords:

subject

Has Abstract

pub_date

2002-05-08 00:00:00

pages

17-22

issue

1-3

eissn

0014-5793

issn

1873-3468

pii

S0014579302026212

journal_volume

518

pub_type

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