PMID- 10602363
OWN - NLM
STAT- MEDLINE
DCOM- 20000214
LR  - 20101118
IS  - 1018-4813 (Print)
IS  - 1018-4813 (Linking)
VI  - 7
IP  - 8
DP  - 1999 Dec
TI  - Carbohydrate-deficient glycoprotein syndrome type 1A: expression and
      characterisation of wild type and mutant PMM2 in E. coli.
PG  - 884-8
AB  - We have identified the PMM2 genotypes of 22 unrelated Danish patients with
      carbohydrate-deficient glycoprotein syndrome type 1A: R141H/F119L (18),
      R141H/C192G (1), F119L/F119L (1), F119L/G117R (1) and D223E/T237R (1). The lack
      of patients homozygous for R141H is statistically highly significant, but
      unexplained. In order to investigate the effect of PMM2 mutations on
      phosphomannomutase (PMM2) activity, PMM2-cDNA was cloned into a pET3a vector.
      Following introduction of mutations into PMM2-cDNA by site-specific mutagenesis, 
      wild type and mutant PMM2-cDNA were expressed in E. coli Bl21(DE3) cells, and the
      activity of PMM2 was determined by an enzymatic assay using mannose 1-phosphate
      as substrate. Recombinant R141H, G117R, and T237R PMM2 had no detectable
      catalytic activity, and the F119L PMM2 had 25% of the activity of the wild type. 
      The activity of the C192G and D223E PMM2 was in the normal range, but the
      affinity for their substrate was lower, and the proteins were more sensitive to
      increased temperatures. Each patient has at least one mutation which retains
      residual PMM2 activity. Our results support the hypotheses that a genotype
      conveying residual PMM2 catalytic activity is required for survival, and that
      homozygosity for R141H impairs PMM2 to a degree incompatible with life.
FAU - Kjaergaard, S
AU  - Kjaergaard S
AD  - Department of Clinical Genetics, Rigshospitalet, University Hospital of
      Copenhagen, Denmark. susanne@rh.dk
FAU - Skovby, F
AU  - Skovby F
FAU - Schwartz, M
AU  - Schwartz M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Eur J Hum Genet
JT  - European journal of human genetics : EJHG
JID - 9302235
RN  - 0 (Acyl Carrier Protein)
RN  - 0 (Mannosephosphates)
RN  - 27251-84-9 (mannose 1-phosphate)
RN  - EC 5.4.2.- (Phosphotransferases (Phosphomutases))
RN  - EC 5.4.2.2 (Phosphoglucomutase)
RN  - EC 5.4.2.8 (phosphomannomutase)
SB  - IM
MH  - Acyl Carrier Protein/*genetics
MH  - Congenital Disorders of Glycosylation/*genetics
MH  - Escherichia coli
MH  - Genotype
MH  - Humans
MH  - Mannosephosphates/metabolism
MH  - Mutation, Missense
MH  - Phosphoglucomutase/*genetics
MH  - Phosphotransferases (Phosphomutases)/*genetics
MH  - Temperature
EDAT- 1999/12/22 09:00
MHDA- 2000/02/19 09:00
CRDT- 1999/12/22 09:00
PHST- 1999/12/22 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 1999/12/22 09:00 [entrez]
AID - 10.1038/sj.ejhg.5200398 [doi]
PST - ppublish
SO  - Eur J Hum Genet. 1999 Dec;7(8):884-8. doi: 10.1038/sj.ejhg.5200398.