PMID- 10387075
OWN - NLM
STAT- MEDLINE
DCOM- 19990722
LR  - 20131121
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 26
DP  - 1999 Jun 29
TI  - The HELLGH motif of rat liver dipeptidyl peptidase III is involved in zinc
      coordination and the catalytic activity of the enzyme.
PG  - 8299-303
AB  - The role of the HELLGH (residues 450-455) motif in the sequence of rat dipeptidyl
      peptidase III (EC 3.4.14.4) was investigated by replacing Glu451 with an alanine 
      or an aspartic acid residue and by replacing His450 and His455 with a tyrosine
      residue by site-directed mutagenesis. Mutated cDNAs were expressed three or four 
      times in Escherichia coli, and the resulting proteins were purified to apparent
      homogeneity. None of the expressed mutated proteins exhibited DPP III activity.
      The mutants of Glu451 contained 1 mol of zinc per mole of protein, but mutants
      His450 and His455 did not contain significant amounts of zinc as determined by
      atomic absorption spectrometry. The Leu453-deleted enzyme (having the zinc
      aminopeptidase motif HExxH-18-E) had almost the same order of binding affinity
      (for Arg-Arg-2-naphthylamide) as the wild-type enzyme, but the specificity
      constant was about 10%. These results provide evidence that the suitable number
      of amino acids included between Glu451 and His455 is three residues for the
      enzyme activity and confirm that residues His450, His455, and Glu451 are involved
      in zinc coordination and catalytic activity.
FAU - Fukasawa, K
AU  - Fukasawa K
AD  - Department of Oral Biochemistry, School of Dentistry, Matsumoto Dental
      University, Shiojiri, Nagano 399-0781, Japan. kmf@po.mdu.ac.jp
FAU - Fukasawa, K M
AU  - Fukasawa KM
FAU - Iwamoto, H
AU  - Iwamoto H
FAU - Hirose, J
AU  - Hirose J
FAU - Harada, M
AU  - Harada M
LA  - eng
SI  - GENBANK/AB017970
SI  - GENBANK/D89340
PT  - Journal Article
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (DNA, Complementary)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 3KX376GY7L (Glutamic Acid)
RN  - 4QD397987E (Histidine)
RN  - EC 3.4.14.- (Dipeptidyl-Peptidases and Tripeptidyl-Peptidases)
RN  - EC 3.4.14.4 (dipeptidyl peptidase III)
RN  - GMW67QNF9C (Leucine)
RN  - J41CSQ7QDS (Zinc)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Catalysis
MH  - DNA, Complementary/isolation & purification
MH  - Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/chemistry/genetics/isolation &
      purification/*physiology
MH  - Enzyme Activation/genetics
MH  - Escherichia coli/genetics
MH  - Glutamic Acid/genetics
MH  - Histidine/genetics
MH  - Humans
MH  - Leucine/genetics
MH  - Liver/*enzymology
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Peptide Fragments/chemistry/genetics/*physiology
MH  - Rats
MH  - Recombinant Proteins/biosynthesis/isolation & purification
MH  - Zinc/chemistry/*metabolism
EDAT- 1999/07/01 00:00
MHDA- 1999/07/01 00:01
CRDT- 1999/07/01 00:00
PHST- 1999/07/01 00:00 [pubmed]
PHST- 1999/07/01 00:01 [medline]
PHST- 1999/07/01 00:00 [entrez]
AID - 10.1021/bi9904959 [doi]
AID - bi9904959 [pii]
PST - ppublish
SO  - Biochemistry. 1999 Jun 29;38(26):8299-303. doi: 10.1021/bi9904959.