PMID- 10029517
OWN - NLM
STAT- MEDLINE
DCOM- 19990316
LR  - 20141120
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 8
DP  - 1999 Feb 23
TI  - Structural elements and limited proteolysis of CD39 influence ATP 
      diphosphohydrolase activity.
PG  - 2248-58
AB  - CD39, the mammalian ATP diphosphohydrolase (ATPDase), is thought to contain two 
      transmembrane domains and five "apyrase conserved regions" (ACR) within a large 
      extracellular region. To study the structure of this ectoenzyme, human CD39 was 
      modified by directed mutations within these ACRs or by sequential deletions at 
      both termini. ATPDase activity was well preserved with FLAG tagging, followed by 
      the removal of either of the demonstrated C- or N-transmembrane regions. However, 
      deletions within ACR-1 (aa 54-61) or -4 (aa 212-220), as well as truncation 
      mutants that included ACR-1, -4, or -5 (aa 447-454), resulted in substantive loss 
      of biochemical activity. Intact ACR-1, -4, and -5 within CD39 are therefore 
      required for maintenance of biochemical activity. Native and mutant forms of CD39 
      lacking TMR were observed to undergo multimerization, associated with the 
      formation of intermolecular disulfide bonds. Limited tryptic cleavage of intact 
      CD39 resulted in two noncovalently membrane-associated fragments (56 and 27 kDa) 
      that substantially augmented ATPDase activity. Glycosylation variation accounted 
      for minor heterogeneity in native and mutant forms of CD39 but did not influence 
      ATPDase function. Enzymatic activity of ATPDase may be influenced by certain 
      posttranslational modifications that are relevant to vascular inflammation.
FAU - Schulte am Esch, J 2nd
AU  - Schulte am Esch J 2nd
AD  - Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical 
      School, Boston, Massachusetts 02215, USA.
FAU - Sevigny, J
AU  - Sevigny J
FAU - Kaczmarek, E
AU  - Kaczmarek E
FAU - Siegel, J B
AU  - Siegel JB
FAU - Imai, M
AU  - Imai M
FAU - Koziak, K
AU  - Koziak K
FAU - Beaudoin, A R
AU  - Beaudoin AR
FAU - Robson, S C
AU  - Robson SC
LA  - eng
GR  - R01HL57307/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Antigens, CD)
RN  - 0 (Isoenzymes)
RN  - 0 (Oligopeptides)
RN  - 0 (Peptide Fragments)
RN  - 0 (Peptides)
RN  - 98849-88-8 (FLAG peptide)
RN  - EC 3.4.- (Endopeptidases)
RN  - EC 3.6.1.- (Adenosine Triphosphatases)
RN  - EC 3.6.1.5 (Apyrase)
RN  - EC 3.6.1.5 (CD39 antigen)
SB  - IM
MH  - *Adenosine Triphosphatases
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antigens, CD/*chemistry/genetics/*metabolism/physiology
MH  - Apyrase/*chemistry/genetics/*metabolism
MH  - Base Sequence
MH  - Blotting, Western
MH  - Cells, Cultured
MH  - Endopeptidases/*metabolism
MH  - Endothelium, Vascular
MH  - Enzyme Activation/genetics
MH  - Flow Cytometry
MH  - Humans
MH  - Hydrolysis
MH  - Isoenzymes/chemistry/genetics/metabolism
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Mutagenesis, Site-Directed
MH  - Oligopeptides
MH  - Peptide Fragments/chemistry/genetics
MH  - Peptides/genetics
MH  - Protein Engineering
MH  - Sequence Deletion
MH  - Swine
MH  - Umbilical Veins
EDAT- 1999/02/25 03:03
MHDA- 2001/03/28 10:01
CRDT- 1999/02/25 03:03
PHST- 1999/02/25 03:03 [pubmed]
PHST- 2001/03/28 10:01 [medline]
PHST- 1999/02/25 03:03 [entrez]
AID - bi982426k [pii]
AID - 10.1021/bi982426k [doi]
PST - ppublish
SO  - Biochemistry. 1999 Feb 23;38(8):2248-58. doi: 10.1021/bi982426k.