Structural effects of naturally occurring human blood group B galactosyltransferase mutations adjacent to the DXD motif
Persson, Mattias ; Letts, James A. ; Hosseini Maaf, Bahram LU ; Borisova, Svetlana N. ; Palcic, Monica M. ; Evans, Stephen V. and Olsson, Martin L LU
(2007)
In Journal of Biological Chemistry
282(13).
p.9564-9570
- Abstract
- Human blood group A and B antigens are produced by two closely related glycosyltransferase enzymes. An N-acetylgalactosaminyltransferase (GTA) utilizes UDP-GalNAc to extend H antigen acceptors (Fuc alpha(1-2)Gal beta-OR) producing A antigens, whereas a galactosyltransferase (GTB) utilizes UDP-Gal as a donor to extend H structures producing B antigens. GTA and GTB have a characteristic (DVD213)-D-211 motif that coordinates to a Mn2+ ion shown to be critical in donor binding and catalysis. Three GTB mutants, M214V, M214T, and M214R, with alterations adjacent to the (211)DVD213 motif have been identified in blood banking laboratories. From serological phenotyping, individuals with the M214R mutation show the B,1 variant expressing very low... (More)
- Human blood group A and B antigens are produced by two closely related glycosyltransferase enzymes. An N-acetylgalactosaminyltransferase (GTA) utilizes UDP-GalNAc to extend H antigen acceptors (Fuc alpha(1-2)Gal beta-OR) producing A antigens, whereas a galactosyltransferase (GTB) utilizes UDP-Gal as a donor to extend H structures producing B antigens. GTA and GTB have a characteristic (DVD213)-D-211 motif that coordinates to a Mn2+ ion shown to be critical in donor binding and catalysis. Three GTB mutants, M214V, M214T, and M214R, with alterations adjacent to the (211)DVD213 motif have been identified in blood banking laboratories. From serological phenotyping, individuals with the M214R mutation show the B,1 variant expressing very low levels of B antigens, whereas those with M214T and M214V mutations give rise to A(weak)B phenotypes. Kinetic analysis of recombinant mutant GTB enzymes revealed that M214R has a 1200-fold decrease in k(cat) compared with wild type GTB. The crystal structure of M214R showed that DVD motif coordination to Mn2+ was disrupted by Arg-214 causing displacement of the metal by a water molecule. Kinetic characterizations of the M214T and M214V mutants revealed they both had GTA and GTB activity consistent with the serology. The crystal structure of the M214T mutant showed no change in DVD coordination to Mn2+. Instead a critical residue, Met-266, which is responsible for determining donor specificity, had adopted alternate conformations. The conformation with the highest occupancy opens up the active site to accommodate the larger A-specific donor, UDP-GalNAc, accounting for the dual specificity. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/667524
- author
- Persson, Mattias
; Letts, James A.
; Hosseini Maaf, Bahram
LU
; Borisova, Svetlana N.
; Palcic, Monica M.
; Evans, Stephen V.
and Olsson, Martin L
LU
- organization
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 282
- issue
- 13
- pages
- 9564 - 9570
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000245421700029
- scopus:34248229370
- pmid:17259183
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M610998200
- language
- English
- LU publication?
- yes
- id
- 3685dec4-19f3-4fc0-a0bd-11d52d0846c2 (old id 667524)
- date added to LUP
- 2016-04-01 12:04:21
- date last changed
- 2022-01-26 22:24:03
@article{3685dec4-19f3-4fc0-a0bd-11d52d0846c2,
abstract = {{Human blood group A and B antigens are produced by two closely related glycosyltransferase enzymes. An N-acetylgalactosaminyltransferase (GTA) utilizes UDP-GalNAc to extend H antigen acceptors (Fuc alpha(1-2)Gal beta-OR) producing A antigens, whereas a galactosyltransferase (GTB) utilizes UDP-Gal as a donor to extend H structures producing B antigens. GTA and GTB have a characteristic (DVD213)-D-211 motif that coordinates to a Mn2+ ion shown to be critical in donor binding and catalysis. Three GTB mutants, M214V, M214T, and M214R, with alterations adjacent to the (211)DVD213 motif have been identified in blood banking laboratories. From serological phenotyping, individuals with the M214R mutation show the B,1 variant expressing very low levels of B antigens, whereas those with M214T and M214V mutations give rise to A(weak)B phenotypes. Kinetic analysis of recombinant mutant GTB enzymes revealed that M214R has a 1200-fold decrease in k(cat) compared with wild type GTB. The crystal structure of M214R showed that DVD motif coordination to Mn2+ was disrupted by Arg-214 causing displacement of the metal by a water molecule. Kinetic characterizations of the M214T and M214V mutants revealed they both had GTA and GTB activity consistent with the serology. The crystal structure of the M214T mutant showed no change in DVD coordination to Mn2+. Instead a critical residue, Met-266, which is responsible for determining donor specificity, had adopted alternate conformations. The conformation with the highest occupancy opens up the active site to accommodate the larger A-specific donor, UDP-GalNAc, accounting for the dual specificity.}},
author = {{Persson, Mattias and Letts, James A. and Hosseini Maaf, Bahram and Borisova, Svetlana N. and Palcic, Monica M. and Evans, Stephen V. and Olsson, Martin L}},
issn = {{1083-351X}},
language = {{eng}},
number = {{13}},
pages = {{9564--9570}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Structural effects of naturally occurring human blood group B galactosyltransferase mutations adjacent to the DXD motif}},
url = {{http://dx.doi.org/10.1074/jbc.M610998200}},
doi = {{10.1074/jbc.M610998200}},
volume = {{282}},
year = {{2007}},
}