Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Ehlers, Mario R.; Fox, Edward A.; Strydom, Daniël J.; Riordan, James

doi:10.1073/pnas.86.20.7741

Cited by 281 publications

(170 citation statements)

References 32 publications

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“…To date, it is difficult to understand the putative functional advantages derived from the complex polyprotein designs occurring in both polyserases as well as in two unrelated human metalloproteases, such as angiotensin-converting enzyme (27)(28)(29) and carboxypeptidase D (30), that also contain different protease domains in their amino acid sequences. In this work, and as a very preliminary attempt to elucidate the physiological functions of the newly identified polyserase-2, we have examined the expression pattern of polyserase-2 in human tissues and tumor cell lines.…”

Section: Discussionmentioning

confidence: 99%

Human Polyserase-2, a Novel Enzyme with Three Tandem Serine Protease Domains in a Single Polypeptide Chain

Cal

Quesada

Llamazares

et al. 2005

Journal of Biological Chemistry

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We have cloned a human cDNA encoding a new serine protease that has been called polyserase-2 (polyserine protease-2) because it is the second identified human enzyme with several tandem serine protease domains in its amino acid sequence. The first serine protease domain contains all characteristic features of these enzymes, whereas the second and third domains lack one residue of the catalytic triad of serine proteases and are predicted to be catalytically inactive. This complex domain organization is also present in the sequences of mouse and rat polyserase-2 and resembles that of polyserase-1, which also contains three serine protease domains in its amino acid sequence. However, polyserase-2 lacks additional domains present in polyserase-1, including a type II transmembrane motif and a low-density lipoprotein receptor A module. Enzymatic analysis demonstrated that both full-length polyserase-2 and its first serine protease domain hydrolyzed synthetic peptides used for assaying serine proteases. Nevertheless, the activity of the isolated domain was greater than that of the entire protein, suggesting that the two catalytically inactive serine protease domains of polyserase-2 may modulate the activity of the first domain. Northern blot analysis showed that polyserase-2 is expressed in fetal kidney; adult skeletal muscle, liver, placenta, prostate, and heart; and tumor cell lines derived from lung and colon adenocarcinomas. Finally, analysis of posttranslational processing mechanisms of polyserase-2 revealed that, contrary to those affecting to the membrane-bound polyserase-1, this novel polyprotein is a secreted enzyme whose three protease domains remain as an integral part of a single polypeptide chain.

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Section: Discussionmentioning

confidence: 99%

Human Polyserase-2, a Novel Enzyme with Three Tandem Serine Protease Domains in a Single Polypeptide Chain

Cal

Quesada

Llamazares

et al. 2005

Journal of Biological Chemistry

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“…Purified human kidney ACE was digested with trypsin, fractionated by HPLC, and the peptides were sequenced by automated Edman degradation as described previously (13).…”

Section: Methodsmentioning

confidence: 99%

Identification of N-Linked Glycosylation Sites in Human Testis Angiotensin-converting Enzyme and Expression of an Active Deglycosylated Form

Sturrock

et al. 1997

Journal of Biological Chemistry

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The sites of glycosylation of Chinese hamster ovary cell expressed testicular angiotensin-converting enzyme (tACE) have been determined by matrix-assisted laser desorption ionization/time of flight/mass spectrometry of peptides generated by proteolytic and cyanogen bromide digestion. Two of the seven potential Nlinked glycosylation sites, Asn 90 and Asn 109 , were found to be fully glycosylated by analysis of peptides before and after treatment with a series of glycosidases and with endoproteinase Asp-N. The mass spectra of the glycopeptides exhibit characteristic clusters of peaks which indicate the N-linked glycans in tACE to be mostly of the biantennary, fucosylated complex type. This structural information was used to demonstrate that three other sites, Asn 155 , Asn 337 , and Asn 586 , are partially glycosylated, whereas Asn 72 appears to be fully glycosylated. The only potential site that was not modified is Asn 620 . Sequence analysis of tryptic peptides obtained from somatic ACE (human kidney) identified six glycosylated and one unglycosylated Asn. Only one of these glycosylation sites had a counterpart in tACE. Comparison of the two proteins reveals a pattern in which amino-terminal N-linked sites are preferred. The functional significance of glycosylation was examined with a tACE mutant lacking the O-glycan-rich first amino-terminal 36 residues and truncated at Ser 625 . When expressed in the presence of the ␣-glucosidase I inhibitor N-butyldeoxynojirimycin and treated with endoglycosidase H to remove all but the terminal N-acetylglucosamine residues, it retained full enzymatic activity, was electrophoretically homogeneous, and is a good candidate for crystallographic studies.Both forms of angiotensin-converting enzyme (ACE 1 ; EC 3.4.15.1 peptidyl-dipeptidase A) are class I transmembrane ectoenzymes (1) that have N-and O-linked oligosaccharides attached to their polypeptide chains (2, 3). Expression of ACE in human HeLa cells in the presence of tunicamycin resulted in complete inhibition of glycosylation, rapidly degraded intracellular ACE, and no enzyme released in the medium (4). An enzymatically active ACE was produced with partial glycosylation in a mutant Chinese hamster ovary (CHO) cell line (ldlD), although it was released to a lesser extent (4). Similarly, it was reported (5) that inhibitors of glucosidases I and II in the endoplasmic reticulum (ER) and mannosidase I in the cis-Golgi reduced the amount of oligosaccharide attached to human intestinal ACE and delayed protein release significantly. These data strongly suggest that glycosylation plays an important role in the membrane targeting and release of ACE, possibly by affecting the folding of the polypeptide and its recognition by a variety of enzymes in the folding and transport machineries. Recently, Sadhukhan and Sen (6) reported that mutations at individual N-linked glycosylation sites (sequons) in rabbit testis ACE (tACE) resulted in varied efficiencies in enzyme release, which suggests that N-linked glycans at each site may make diff...

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“…The human ACE gene is located in chromosome 17q23 with 26 exons and 25 introns with an insertion/deletion (I/D) polymorphism that comprises a 278-bp fragment in intron 16 (3). The DD genotype or D allele of this polymorphism was shown to be associated with elevated circulating and tissue ACE activity (4) as well as increased risk of hypertension (5) and diabetic renal (6) and cardiovascular complications (7).…”

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confidence: 99%

Prognostic Effect of Insertion/Deletion Polymorphism of the ACE Gene on Renal and Cardiovascular Clinical Outcomes in Chinese Patients With Type 2 Diabetes

Wang

et al. 2005

Diabetes Care

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OBJECTIVE -The insertion/deletion (I/D) polymorphism of the ACE gene has been reported to be associated with diabetic microvascular or macrovascular complications. The aim of the present study was to investigate the prognostic effect of I/D polymorphism on renal and cardiovascular clinical outcomes in Chinese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS -A consecutive cohort of 1,281Chinese patients with type 2 diabetes were followed for 41.3 Ϯ 21.6 months. Renal end points were defined as renal death and events (need for dialysis, plasma creatinine Ն500 mol/l, or doubling of plasma creatinine of baseline value Ն150 mol/l). Cardiovascular end points were defined as cardiovascular death and events, which included ischemic heart disease, heart failure, cerebrovascular accident, and revascularization requiring hospital admission. The I/D polymorphism of the ACE gene was examined by PCR followed by agarose gel electrophoresis.RESULTS -The frequencies of ACE gene I/D polymorphisms were in Hardy-Weinberg equilibrium. Patients who developed a renal end point (n ϭ 98) had higher frequencies of DD genotype (19.4 vs. 10.8%, P ϭ 0.018) and D allele (41.3 vs. 31.8%, P ϭ 0.006) compared with subjects who did not (n ϭ 1,183). The cumulative rates of renal end points were 10.0, 19.2, and 24.4% in the II (n ϭ 595), DI (n ϭ 539), and DD genotype carriers (n ϭ 147), respectively (log rank P ϭ 0.004). In multiple Cox regression analysis, the occurrence of renal end points remained significantly influenced by I/D polymorphism with a dominant deleterious effect of the DD genotype (DD versus II, adjusted hazard ratio 2.80 [95% CI 1.49 -5.29]). There was no prognostic effect of I/D polymorphism on cardiovascular end points.CONCLUSIONS -The DD genotype of the ACE I/D polymorphism was an independent risk factor for renal but not cardiovascular end points in Chinese patients with type 2 diabetes. Diabetes Care 28:348 -354, 2005A CE is one of the key enzymes in the renin-angiotensin system, which plays an important role in fluid and electrolyte balance and regulation of blood pressure and cellular growth (1,2). The human ACE gene is located in chromosome 17q23 with 26 exons and 25 introns with an insertion/deletion (I/D) polymorphism that comprises a 278-bp fragment in intron 16 (3). The DD genotype or D allele of this polymorphism was shown to be associated with elevated circulating and tissue ACE activity (4) as well as increased risk of hypertension (5) and diabetic renal (6) and cardiovascular complications (7). However, these results remained inconsistent in both Caucasian (8 -11) and non-Caucasian populations, including Chinese (12,13). These discrepancies might be due to ethnicity, study design, patient selection criteria, and small sample size. The aim of the present study was to evaluate the prognostic effect of this polymorphism on renal and cardiovascular outcomes in a large cohort of 1,281 Chinese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS -The Prince of WalesHospital is the teaching hospital of t...

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Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Cited by 281 publications

References 32 publications

Human Polyserase-2, a Novel Enzyme with Three Tandem Serine Protease Domains in a Single Polypeptide Chain

Human Polyserase-2, a Novel Enzyme with Three Tandem Serine Protease Domains in a Single Polypeptide Chain

Identification of N-Linked Glycosylation Sites in Human Testis Angiotensin-converting Enzyme and Expression of an Active Deglycosylated Form

Prognostic Effect of Insertion/Deletion Polymorphism of the ACE Gene on Renal and Cardiovascular Clinical Outcomes in Chinese Patients With Type 2 Diabetes

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