Purification and characterization of human metallocarboxypeptidase Z

Elena G. Novikova, Lloyd D. Fricker

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Carboxypeptidase Z (CPZ) is a recently discovered member of the metallocarboxypeptidase gene family that has an N-terminal domain related to the Wnt/wingless binding domain of frizzled receptors and other proteins. To further characterize the enzymatic properties of CPZ, the enzyme was purified using Arg-and heparin-affinity columns. CPZ has a neutral pH optimum, and is inhibited by chelating agents and several divalent cations (Zn2+, Mn2+, Cd2+, Cu2+, Hg2+). Active site-directed inhibitors of several other metallocarboxypeptidases also inhibit CPZ activity with moderate potency. CPZ cleaves substrates with C-terminal Arg residues, preferring peptides with an Ala in the penultimate position. No activity is detected toward substrates with an Ile-Arg or a Pro-Arg sequence. The Km for dansyl-Phe-Ala-Arg and dansyl-Pro-Ala-Arg are both approximately 2 mM. Taken together, these data suggests a selective role for CPZ in the processing of extracellular peptides or proteins.

Original languageEnglish (US)
Pages (from-to)564-568
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume256
Issue number3
DOIs
StatePublished - Mar 24 1999

Fingerprint

Purification
prolylarginine
Frizzled Receptors
Peptides
Divalent Cations
Substrates
Chelating Agents
Heparin
metallocarboxypeptidase Z
carboxypeptidase Z
Catalytic Domain
Proteins
Genes
Enzymes
Processing

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification and characterization of human metallocarboxypeptidase Z. / Novikova, Elena G.; Fricker, Lloyd D.

In: Biochemical and Biophysical Research Communications, Vol. 256, No. 3, 24.03.1999, p. 564-568.

Research output: Contribution to journalArticle

@article{7b8b1f10092e4075b1402400ca18a50b,
title = "Purification and characterization of human metallocarboxypeptidase Z",
abstract = "Carboxypeptidase Z (CPZ) is a recently discovered member of the metallocarboxypeptidase gene family that has an N-terminal domain related to the Wnt/wingless binding domain of frizzled receptors and other proteins. To further characterize the enzymatic properties of CPZ, the enzyme was purified using Arg-and heparin-affinity columns. CPZ has a neutral pH optimum, and is inhibited by chelating agents and several divalent cations (Zn2+, Mn2+, Cd2+, Cu2+, Hg2+). Active site-directed inhibitors of several other metallocarboxypeptidases also inhibit CPZ activity with moderate potency. CPZ cleaves substrates with C-terminal Arg residues, preferring peptides with an Ala in the penultimate position. No activity is detected toward substrates with an Ile-Arg or a Pro-Arg sequence. The Km for dansyl-Phe-Ala-Arg and dansyl-Pro-Ala-Arg are both approximately 2 mM. Taken together, these data suggests a selective role for CPZ in the processing of extracellular peptides or proteins.",
author = "Novikova, {Elena G.} and Fricker, {Lloyd D.}",
year = "1999",
month = "3",
day = "24",
doi = "10.1006/bbrc.1999.0378",
language = "English (US)",
volume = "256",
pages = "564--568",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Purification and characterization of human metallocarboxypeptidase Z

AU - Novikova, Elena G.

AU - Fricker, Lloyd D.

PY - 1999/3/24

Y1 - 1999/3/24

N2 - Carboxypeptidase Z (CPZ) is a recently discovered member of the metallocarboxypeptidase gene family that has an N-terminal domain related to the Wnt/wingless binding domain of frizzled receptors and other proteins. To further characterize the enzymatic properties of CPZ, the enzyme was purified using Arg-and heparin-affinity columns. CPZ has a neutral pH optimum, and is inhibited by chelating agents and several divalent cations (Zn2+, Mn2+, Cd2+, Cu2+, Hg2+). Active site-directed inhibitors of several other metallocarboxypeptidases also inhibit CPZ activity with moderate potency. CPZ cleaves substrates with C-terminal Arg residues, preferring peptides with an Ala in the penultimate position. No activity is detected toward substrates with an Ile-Arg or a Pro-Arg sequence. The Km for dansyl-Phe-Ala-Arg and dansyl-Pro-Ala-Arg are both approximately 2 mM. Taken together, these data suggests a selective role for CPZ in the processing of extracellular peptides or proteins.

AB - Carboxypeptidase Z (CPZ) is a recently discovered member of the metallocarboxypeptidase gene family that has an N-terminal domain related to the Wnt/wingless binding domain of frizzled receptors and other proteins. To further characterize the enzymatic properties of CPZ, the enzyme was purified using Arg-and heparin-affinity columns. CPZ has a neutral pH optimum, and is inhibited by chelating agents and several divalent cations (Zn2+, Mn2+, Cd2+, Cu2+, Hg2+). Active site-directed inhibitors of several other metallocarboxypeptidases also inhibit CPZ activity with moderate potency. CPZ cleaves substrates with C-terminal Arg residues, preferring peptides with an Ala in the penultimate position. No activity is detected toward substrates with an Ile-Arg or a Pro-Arg sequence. The Km for dansyl-Phe-Ala-Arg and dansyl-Pro-Ala-Arg are both approximately 2 mM. Taken together, these data suggests a selective role for CPZ in the processing of extracellular peptides or proteins.

UR - http://www.scopus.com/inward/record.url?scp=0033599517&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033599517&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1999.0378

DO - 10.1006/bbrc.1999.0378

M3 - Article

C2 - 10080937

AN - SCOPUS:0033599517

VL - 256

SP - 564

EP - 568

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 3

ER -