TY - JOUR

T1 - Peptidyl dipeptidases (Ance and Acer) of Drosophila melanogaster: major differences in the substrate specificity of two homologs of human angiotensin I-converting enzyme.

AU - Siviter, Richard J.

AU - Nachman, Ronald J.

AU - Dani, M. Paulina

AU - Keen, Jeffrey N.

AU - Shirras, Alan D.

AU - Isaac, R. Elwyn

PY - 2002/11

Y1 - 2002/11

N2 - Drosophila melanogaster angiotensin converting enzyme (Ance) and angiotensin converting enzyme related (Acer) are single domain homologs of mammalian peptidyl dipeptidase A (angiotensin I-converting enzyme) whose physiological substrates have not as yet been identified. We have investigated the in vitro substrate specificities of the two peptidases towards a variety of insect and mammalian peptides. Ance was generally much better than Acer at hydrolyzing peptides of 5–13 amino acids in length. Only two of the peptides, [Leu5]enkephalinamide and leucokinin-I were cleaved faster by Acer. Increasing NaCl concentration had opposite affects on the cleavage of [Leu5]enkephalin and [Leu5]enkephalinamide by Acer, decreasing the activity towards [Leu5]enkephalin but increasing the activity towards [Leu5]enkephalinamide. Of the insect peptides tested, the tachykinin-related peptide, Lom TK-1, proved to be the best substrate for Ance with a kcat/Km ratio of 0.122 s−1 μM−1. However, in comparison, the D. melanogaster tachykinins, DTK-1, DTK-2, DTK-3 and DTK-4 were poor Ance substrates. DTK-5 was the best substrate of this family, but the apparent high Km for hydrolysis by Ance suggested that this peptide would not be a natural Ance substrate. This low affinity for DTK-5 is the likely reason why the peptide was not rapidly degraded in D. melanogaster hemolymph, where Ance was shown to be a major peptide-degrading activity.

AB - Drosophila melanogaster angiotensin converting enzyme (Ance) and angiotensin converting enzyme related (Acer) are single domain homologs of mammalian peptidyl dipeptidase A (angiotensin I-converting enzyme) whose physiological substrates have not as yet been identified. We have investigated the in vitro substrate specificities of the two peptidases towards a variety of insect and mammalian peptides. Ance was generally much better than Acer at hydrolyzing peptides of 5–13 amino acids in length. Only two of the peptides, [Leu5]enkephalinamide and leucokinin-I were cleaved faster by Acer. Increasing NaCl concentration had opposite affects on the cleavage of [Leu5]enkephalin and [Leu5]enkephalinamide by Acer, decreasing the activity towards [Leu5]enkephalin but increasing the activity towards [Leu5]enkephalinamide. Of the insect peptides tested, the tachykinin-related peptide, Lom TK-1, proved to be the best substrate for Ance with a kcat/Km ratio of 0.122 s−1 μM−1. However, in comparison, the D. melanogaster tachykinins, DTK-1, DTK-2, DTK-3 and DTK-4 were poor Ance substrates. DTK-5 was the best substrate of this family, but the apparent high Km for hydrolysis by Ance suggested that this peptide would not be a natural Ance substrate. This low affinity for DTK-5 is the likely reason why the peptide was not rapidly degraded in D. melanogaster hemolymph, where Ance was shown to be a major peptide-degrading activity.

KW - Insect tachykinin

KW - Insect peptides

KW - Peptide metabolism

KW - Peptidases

KW - Tachykinin-related peptides

U2 - 10.1016/S0196-9781(02)00190-0

DO - 10.1016/S0196-9781(02)00190-0

M3 - Journal article

VL - 23

SP - 2025

EP - 2034

JO - Peptides

JF - Peptides

SN - 0196-9781

IS - 11

ER -