PMID- 2250024
OWN - NLM
STAT- MEDLINE
DCOM- 19910117
LR  - 20220309
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 265
IP  - 34
DP  - 1990 Dec 5
TI  - Rat brain N-acetylated alpha-linked acidic dipeptidase activity. Purification and 
      immunologic characterization.
PG  - 21297-301
AB  - N-Acetylated alpha-linked acidic dipeptidase (NAALA dipeptidase) is a 
      membrane-bound metallopeptidase that cleaves glutamate from the endogenous 
      neuropeptide N-acetyl-L-aspartyl-L-glutamate. In this report, we have solubilized 
      NAALA dipeptidase activity from synaptosomal membranes with Triton X-100 and 
      purified it to apparent homogeneity by sequential column chromatography on 
      DEAE-Sepharose, CM-Sepharose, and lentil lectin-Sepharose. This procedure 
      resulted in a 720-fold purification with 1.6% yield. The purified ezyme migrated 
      as a single silver-stained band on a sodium dodecyl sulfate gel with an apparent 
      molecular weight of 94 kDa. Using an enzymatic stain to visualize NAALA 
      dipeptidase activity within a gel matrix, we have confirmed that the 94-kDa band 
      is, indeed, NAALA dipeptidase. The purified enzyme was characterized and found to 
      be pharmacologically similar to NAALA dipeptidase activity described previously 
      in synaptosomal membrane extracts. Using the purified NAALA dipeptidase as 
      antigen, we have raised specific and high titer polyclonal antibodies in guinea 
      pig. Immunocytochemical studies show intense NAALA dipeptidase immunoreactivity 
      in the cerebellar and renal cortices.
FAU - Slusher, B S
AU  - Slusher BS
AD  - Department of Neuroscience and Pharmacology, Johns Hopkins School of Medicine, 
      Baltimore, Maryland 21205.
FAU - Robinson, M B
AU  - Robinson MB
FAU - Tsai, G
AU  - Tsai G
FAU - Simmons, M L
AU  - Simmons ML
FAU - Richards, S S
AU  - Richards SS
FAU - Coyle, J T
AU  - Coyle JT
LA  - eng
GR  - 2 T32 GM07626/GM/NIGMS NIH HHS/United States
GR  - NS 07870/NS/NINDS NIH HHS/United States
GR  - NS 13584/NS/NINDS 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  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antigen-Antibody Complex)
RN  - EC 3.4.13.- (Dipeptidases)
RN  - EC 3.4.17.21 (Glutamate Carboxypeptidase II)
SB  - IM
MH  - Animals
MH  - Antigen-Antibody Complex/analysis
MH  - Brain/*enzymology
MH  - Chromatography, Affinity
MH  - Chromatography, Gel
MH  - Chromatography, Ion Exchange
MH  - Dipeptidases/immunology/isolation & purification/*metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Glutamate Carboxypeptidase II
MH  - Intracellular Membranes/enzymology
MH  - Kinetics
MH  - Male
MH  - Molecular Weight
MH  - Organ Specificity
MH  - Rats
MH  - Rats, Inbred Strains
MH  - Synaptosomes/enzymology
EDAT- 1990/12/05 00:00
MHDA- 1990/12/05 00:01
CRDT- 1990/12/05 00:00
PHST- 1990/12/05 00:00 [pubmed]
PHST- 1990/12/05 00:01 [medline]
PHST- 1990/12/05 00:00 [entrez]
AID - S0021-9258(17)45359-2 [pii]
PST - ppublish
SO  - J Biol Chem. 1990 Dec 5;265(34):21297-301.