Studies on Procarboxypeptidase a and Carboxypeptidase a of the Spiny Pacific Dogfish (Squalus acanthias)

Andras G. Lacko, Hans Neurath

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

A procedure is described for the purification of procarboxypeptidase A from acetone powder prepared from extracts of pancreas glands of the spiny Pacific dogfish. The apparently homogeneous zymogen is a monomer in contrast to the procarboxypeptidase A complex isolated from the bovine pancreas. Physical and chemical properties of dogfish procarboxypeptidase A were found to be similar to those of succinyl fraction I, the chemically modified, monomeric precursor of bovine carboxypeptidase A. Significant differences, however, exist in the number of sulfur-containing residues in each zymogen. Dogfish procarboxypeptidase A was rapidly activated by bovine trypsin or Nagarse in the presence of Ca2+ ions (up to 0.5 m). Activation by chymotrypsin resulted in the generation of only about 18% of the total potential activity of the zymogen. Calcium ions increased neither the efficiency nor the extent of activation by chymotrypsin. Dogfish carboxypeptidase A was isolated following the activation of purified procarboxypeptidase A with bovine trypsin. The enzyme appeared to be homogeneous in the ultracentrifuge and resembled its bovine counterpart in physical, chemical, and catalytic properties. Dogfish procarboxypeptidase A was also found to have catalytic activity toward ester and peptide substrates. Comparison of the catalytic properties of the enzyme and the zymogen reveals that the product of the activation process probably has a more efficient binding site rather than a more effective catalytic site.

Original languageEnglish
Pages (from-to)4680-4690
Number of pages11
JournalBiochemistry
Volume9
Issue number24
DOIs
StatePublished - 1 Nov 1970

Fingerprint

Squalus acanthias
Carboxypeptidases A
Carboxypeptidases
Dogfish
Enzyme Precursors
Chemical activation
Chymotrypsin
Trypsin
Pancreas
Ions
Subtilisins
Enzymes
Acetone
Sulfur
Powders
Chemical properties
Purification
Catalyst activity
Catalytic Domain
Esters

Cite this

@article{faf8a4247ec840fa9a4d6df53e5225ec,
title = "Studies on Procarboxypeptidase a and Carboxypeptidase a of the Spiny Pacific Dogfish (Squalus acanthias)",
abstract = "A procedure is described for the purification of procarboxypeptidase A from acetone powder prepared from extracts of pancreas glands of the spiny Pacific dogfish. The apparently homogeneous zymogen is a monomer in contrast to the procarboxypeptidase A complex isolated from the bovine pancreas. Physical and chemical properties of dogfish procarboxypeptidase A were found to be similar to those of succinyl fraction I, the chemically modified, monomeric precursor of bovine carboxypeptidase A. Significant differences, however, exist in the number of sulfur-containing residues in each zymogen. Dogfish procarboxypeptidase A was rapidly activated by bovine trypsin or Nagarse in the presence of Ca2+ ions (up to 0.5 m). Activation by chymotrypsin resulted in the generation of only about 18{\%} of the total potential activity of the zymogen. Calcium ions increased neither the efficiency nor the extent of activation by chymotrypsin. Dogfish carboxypeptidase A was isolated following the activation of purified procarboxypeptidase A with bovine trypsin. The enzyme appeared to be homogeneous in the ultracentrifuge and resembled its bovine counterpart in physical, chemical, and catalytic properties. Dogfish procarboxypeptidase A was also found to have catalytic activity toward ester and peptide substrates. Comparison of the catalytic properties of the enzyme and the zymogen reveals that the product of the activation process probably has a more efficient binding site rather than a more effective catalytic site.",
author = "Lacko, {Andras G.} and Hans Neurath",
year = "1970",
month = "11",
day = "1",
doi = "10.1021/bi00826a010",
language = "English",
volume = "9",
pages = "4680--4690",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "24",

}

Studies on Procarboxypeptidase a and Carboxypeptidase a of the Spiny Pacific Dogfish (Squalus acanthias). / Lacko, Andras G.; Neurath, Hans.

In: Biochemistry, Vol. 9, No. 24, 01.11.1970, p. 4680-4690.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Studies on Procarboxypeptidase a and Carboxypeptidase a of the Spiny Pacific Dogfish (Squalus acanthias)

AU - Lacko, Andras G.

AU - Neurath, Hans

PY - 1970/11/1

Y1 - 1970/11/1

N2 - A procedure is described for the purification of procarboxypeptidase A from acetone powder prepared from extracts of pancreas glands of the spiny Pacific dogfish. The apparently homogeneous zymogen is a monomer in contrast to the procarboxypeptidase A complex isolated from the bovine pancreas. Physical and chemical properties of dogfish procarboxypeptidase A were found to be similar to those of succinyl fraction I, the chemically modified, monomeric precursor of bovine carboxypeptidase A. Significant differences, however, exist in the number of sulfur-containing residues in each zymogen. Dogfish procarboxypeptidase A was rapidly activated by bovine trypsin or Nagarse in the presence of Ca2+ ions (up to 0.5 m). Activation by chymotrypsin resulted in the generation of only about 18% of the total potential activity of the zymogen. Calcium ions increased neither the efficiency nor the extent of activation by chymotrypsin. Dogfish carboxypeptidase A was isolated following the activation of purified procarboxypeptidase A with bovine trypsin. The enzyme appeared to be homogeneous in the ultracentrifuge and resembled its bovine counterpart in physical, chemical, and catalytic properties. Dogfish procarboxypeptidase A was also found to have catalytic activity toward ester and peptide substrates. Comparison of the catalytic properties of the enzyme and the zymogen reveals that the product of the activation process probably has a more efficient binding site rather than a more effective catalytic site.

AB - A procedure is described for the purification of procarboxypeptidase A from acetone powder prepared from extracts of pancreas glands of the spiny Pacific dogfish. The apparently homogeneous zymogen is a monomer in contrast to the procarboxypeptidase A complex isolated from the bovine pancreas. Physical and chemical properties of dogfish procarboxypeptidase A were found to be similar to those of succinyl fraction I, the chemically modified, monomeric precursor of bovine carboxypeptidase A. Significant differences, however, exist in the number of sulfur-containing residues in each zymogen. Dogfish procarboxypeptidase A was rapidly activated by bovine trypsin or Nagarse in the presence of Ca2+ ions (up to 0.5 m). Activation by chymotrypsin resulted in the generation of only about 18% of the total potential activity of the zymogen. Calcium ions increased neither the efficiency nor the extent of activation by chymotrypsin. Dogfish carboxypeptidase A was isolated following the activation of purified procarboxypeptidase A with bovine trypsin. The enzyme appeared to be homogeneous in the ultracentrifuge and resembled its bovine counterpart in physical, chemical, and catalytic properties. Dogfish procarboxypeptidase A was also found to have catalytic activity toward ester and peptide substrates. Comparison of the catalytic properties of the enzyme and the zymogen reveals that the product of the activation process probably has a more efficient binding site rather than a more effective catalytic site.

UR - http://www.scopus.com/inward/record.url?scp=0014961128&partnerID=8YFLogxK

U2 - 10.1021/bi00826a010

DO - 10.1021/bi00826a010

M3 - Article

C2 - 5529813

AN - SCOPUS:0014961128

VL - 9

SP - 4680

EP - 4690

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 24

ER -