PubMed Health⌕ Search

Biomedical subjects

T Vanha-Perttula

Publications and source records attributed to T Vanha-Perttula.

At least 55 records · Page 3Linked to original sources
PubMed1985

Variable distribution of aminopeptidase A in male reproductive organs of mammals.

Aminopeptidase A (AP-A) was analysed in the reproductive organs of the boar, bull, gerbil and man. High hydrolysis of alpha-L-glutamyl-beta-naphthylamide (GluNA) and alpha-L-aspartyl-beta-naphthylamide (AspNA) with activation by alkaline earth metals was detected in the ampulla, seminal vesicles, and seminal vesicle secretions of the bull and in the cauda epididymis of the boar and gerbil. In man, weak AP-A activity was found in all reproductive tissues. Histochemically, AP-A was localized in the epithelial cells of tissues having a high specific activity for the enzyme. AP-A was absent from human seminal fluid, whilst bovine seminal fluid had strong, and boar seminal fluid weaker, AP-A activity. Gel filtration of bull seminal vesicle secretions and seminal fluid, boar seminal fluid or an homogenate of boar and gerbil epididymal cauda and human epididymis and seminal vesicles on Sephacryl S-300 resulted in a major high-molecular-weight activity peak A at Ve/Vo = 1.17 and another low-molecular-weight peak B at Ve/Vo = 1.51 (man), 1.62 (boar, bull) or 1.75 (gerbil). This fractionation was not in all cases able to separate AP-A from aminopeptidase(s), which were active on L-alanine-beta-naphthylamide (AlaNA) but showed no activation by alkaline earth metals. Homogenates of bovine epididymis showed only the low-molecular-weight GluNA peak B, but two areas of activity for AlaNA hydrolysis. In bovine seminal vesicles and porcine epididymis, AP-A activity appeared to be linked with the functional maturity of these organs. The high-molecular-weight AP-A (peak A) appeared to be the predominant form in seminal fluid.

Aminopeptidases↗
PubMed1985

Acid and neutral alpha-glucosidase in the reproductive organs and seminal plasma of the bull.

A synthetic substrate (p-nitrophenyl-alpha-D-glucopyranoside) was used to measure the acid and neutral alpha-glucosidase activity in bull seminal plasma, spermatozoa and in homogenates of bull reproductive organs. Marked differences were observed in the activities of these enzymes in the various tissues studied. Epididymis and particularly its caput region contained the highest specific activity of acid alpha-glucosidase. The activity of neutral alpha-glucosidase was highest in testis and in different parts of the epididymis. Seminal plasma, spermatozoa and seminal vesicle secretion contained only the acid enzyme activity. After fractionation with anion exchange chromatography in HPLC (Mono Q) and chromatofocussing, acid alpha-glucosidase activity of seminal plasma was recovered in two fractions with different pI values. The corresponding activities were found in the secretion of seminal vesicles, which thus form the major secretory source of seminal plasma acid alpha-glucosidase. In the fractionation with gel filtration on Sepharose 6B, the acid alpha-glucosidase had a smaller molecular weight than did the neutral enzyme. In anion exchange chromatography and chromatofocussing the testicular and epididymal homogenates each contained two acid and two neutral isoenzymes. In both fractionations the elution pattern of acid alpha-glucosidase was clearly different from that of the enzymes in seminal plasma. The pH optimum of acid alpha-glucosidase ranged from 3.75 to 4.5 and that of the neutral enzyme from 6.5 to 7.0. The neutral activity was more sensitive to many divalent metal ions and differences were also observed in the response of the enzymes to different concentrations of turanose and KCl.

Animals↗
PubMed1984

Purification and partial characterization of aminopeptidase A from the serum of pregnant and non-pregnant women.

Aminopeptidase A (L-alpha-aspartyl(L-alpha-glutamyl)-peptide hydrolase, EC 3.4.11.7) was purified from human maternal and control sera using CM-cellulose chromatography, DEAE-Sephacel chromatography, Sephacryl S-300 gel filtration and hydroxyapatite chromatography. The purification coefficients were 3069 and 5210 and the yields 6.3 and 6.1% for the maternal and control serum, respectively. The purified enzymes appeared free from other serum aminopeptidases in polyacrylamide gel electrophoresis. The biochemical and physical characteristics of the enzymes from maternal and control sera were similar. A molecular weight of 260 000, an optimum at pH 6.75-7.25 and a fairly good stability of the enzymes at 4 and -18 degrees C were recorded. The alkaline earth metals (Ca2+, Ba2+, Sr2+) were the activators of alpha-L-glutamyl-beta-naphthylamide hydrolysis, while alpha-L-aspartyl-beta-naphthylamide hydrolysis was markedly potentiated with Ca2+ but not with Ba2+ at all. The most rapid hydrolysis was shown with GluNA (Km with Ba2+ 0.156 +/- 0.014 mM and 0.136 +/- 0.009 mM in maternal and control serum, respectively), while only minimal hydrolysis of some neutral and basic amino-acid-beta-naphthylamides were observed. The contribution of the placenta to the elevated aminopeptidase A levels in the pregnancy plasma could not be solved on the basis of the present observations.

Amino Acids↗
PubMed1984

Comparison of acid phosphatases in the rat prostatic complex and seminal vesicles.

Acid phosphatase activity of different parts (ventral, lateral and posterior lobes and coagulating gland) of the rat prostatic complex and seminal vesicles were analyzed in homogenate and after fractionation with gel filtration and chromatofocusing. Significant differences between the various tissue homogenates were recorded in the hydrolysis of p-nitrophenyl phosphate, alpha-naphthyl phosphate and naphthol ASBI phosphate and the percentage of the tartrate-resistant activity varied from about 30 (in seminal vesicles) to 56 (in coagulating gland). Gel filtration resulted in the appearance of 2 (Sephadex G-200) to 3 (Sepharose 6B) activity peaks (GF-1 to GF-3). Studies with 6 substrates (p-nitrophenyl phosphate, alpha-naphthyl phosphate, beta-naphthyl phosphate, naphthol ASBI phosphate, phenolphthalein phosphate and thymolphthalein phosphate) showed some relative differences in the hydrolysis rates as well as indications for possible overlapping of multiple enzymes in these peaks. Chromatofocusing of the ventral prostate homogenate resulted in the appearance of 8 activities (CF-1 to CF-8) with different pI-values (8.2, 8.1, 7.9, 7.1, 6.4, 6.0, 5.5, and 5.0), substrate preferences, pH-optima (5.5, 4.2, 4.2, 6.0, 5.5, 5.0, 4.2, and 5.0), molecular weights and modifier characteristics. The other tissues contained a lower number of these activities: 4 (CF-1 to CF-3 and CF-6) in lateral and posterior lobes and coagulating gland and 5 (CF-1 to CF-4 and CF-6) in seminal vesicles. The correspondence of the enzyme peaks after gel filtration and chromatofocusing was analyzed by refractionation. This indicated that some of the enzymes (GF-1, CF-5, CF-8) may appear in polymeric/aggregate forms. The differences in the acid phosphatase pattern suggest characteristic functional features in the various parts of the rat prostatic complex.

Acid Phosphatase↗
PubMed1984

Aminopeptidase A in reproductive organs of the male rat: evidence for high activity in the posterior lobe of the prostate.

Of the reproductive organs of the male rat a high level of hydrolysis of alpha-L-glutamyl-beta-napthylamide (GluNA) and alpha-L-aspartyl-beta-naphthylamide (AspNA) in the presence of Ca2+ (5mM) (aminopeptidase A) was found in the posterior lobe of the prostate. Histochemically, this enzyme was localized in the epithelial cells of acini, which were grouped in the dorsal part and sharply separated from non-active acini in the anterior part of the lobe. A single peak of Ca2+-activated GluNA and AspNA hydrolysis was obtained after chromatofocusing at pI 4.9 and on anion exchange chromatography this activity eluted at 0.09 M NaCl. After gel filtration on Sepharose 6B a major peak of activity was found at the elution volume (Ve/Vo = 2.28). In all of these fractionation procedures aminopeptidase A was partially or totally overlapped by other aminopeptidases hydrolysing various amino acid-beta-naphthylamides. A pooled enzyme preparation gave an optimum at pH 7.3. The hydrolysis of GluNA was markedly enhanced in the presence of Ba2+, Ca2+ and Sr2+, but the hydrolysis of AspNA was activated only by Ca2+ and Sr2+. Castration caused a significant decrease in the hydrolysis of GluNA by the posterior lobe, but did not influence the low levels of activity in other parts of the prostate or in the seminal vesicles.

Amino Acids↗
PubMed1983

Separation of acid phosphatases in the rat ventral prostate by gel filtration, isoelectric focusing, and chromatofocusing.

Acid phosphatases of the rat ventral prostate were fractionated by gel filtration (GF) on Sepharose 6B, isoelectric focusing (IEF), and chromatofocusing (CF). In GF three activity peaks (GF-1, GF-2, GF-3) were disclosed. They showed some differences in substrate preference when six substrates (p-nitrophenyl phosphate; p-NPP; phenolphthalein phosphate, Phe-P; thymolphthalein phosphate, Tym-P; alpha-naphthyl phosphate, alpha-NP; beta-naphthyl phosphate, beta-NP; naphthol ASBI phosphate, N-ASBI-P) were tested. Differences were also encountered in their sensitivity to tartrate and fluoride. IEF gave seven bands at different pI values (8.3, 8.1, 7.9, 7.1, 6.4, 5.5, and 5.0) with alpha-NP and beta-NP but only four with N-ASBI-P. Four of the bands (8.3, 8.1, 7.9, 5.5) were sensitive to tartrate. In CF eight activity peaks (CF-1 to CF-8) were resolved with the six substrates. They differed from each other in pI values, pH optima, substrate preference, and modifier characteristics. Peaks CF-1 (pI 8.3, pH 5.5), CF-2 (pI 8.1, pH 4.2) and CF-3 (pI 7.9, pH 4.2) had a large substrate spectrum and high sensitivity to tartrate and fluoride. CF-4 (pI 7.1, pH 6.0) and CF-7 (pI 5.5, pH 4.2) were low in activity, preferred alpha-NP as substrate, and were moderately sensitive to tartrate. CF-5 (pI 6.4, pH 5.5) and CF-8 (pI 5.0, pH 5.0) were able to hydrolyse all substrates tested with moderate inhibition by tartrate. CF-6 (pI 6.0, pH 5.0) showed a relative preference for p-NPP and Phe-P with no hydrolysis of N-ASBI-P and Tym-P. Of these activities CF-6 and CF-7 were also clearly activated by Co2+. Peaks CF-6 and CF-7 appeared the most sensitive to p-chloromercuribenzoate. It is concluded that activities CF-1, CF-2, and CF-3 are lysosomal isoenzymes with minor structural differences. The others are possibly all nonlysosomal with greater biochemical differences. Some of them apparently represent the secretory form(s) of acid phosphatase in the rat ventral prostate.

Acid Phosphatase↗
PubMed1983

Characterization of two thiol-dependent aminopeptidases partially purified from human placenta.

1. Two thiol-dependent aminopeptidases (I and III) were partially purified from the soluble fraction of human placenta. 2. Aminopeptidase I preferred L-alanine-beta-naphthylamide (AlaNA) as substrate at pH 7.0-7.5. It was sensitive to heat and some divalent metal ions (Co2+, Ni2+, Zn2+) but resistant to EDTA, and some amino acids. 3. Aminopeptidase III hydrolysed equally well AlaNA and ArgNA at pH 6.5-7.0. It was markedly suppressed by EDTA and reactivated by Co2+. It was inhibited by heat, some amino acids, benzamadine and puromycin.

Aminopeptidases↗
PubMed1983

Aminopeptidases of the rat prostatic complex and seminal vesicles: separation by gel filtration and chromatofocusing.

Aminopeptidases hydrolysing L-alanine-beta-naphthylamide (AlaNA), L-arginine-beta-naphthylamide (ArgNA) and L-leucine-beta-napthylamide (LeuNA) were studied in homogenate and after fractionation (gel filtration, chromatofocusing) of various parts of the rat prostatic complex (ventral lobe, VP; lateral lobe, LP; posterior lobe, PP; coagulating gland, CG) and seminal vesicles (VS). Marked differences were observed in the hydrolysis rates of these substrates in the various tissues studied. After fractionation up to seven activities were separated with differences in pH-optima, pI-values, molecular weights, substrate preferences and modifier characteristics. The number of enzyme activities characteristically varied between the tissues studied. Depending on the enzyme pattern in each part the differences in hydrolysis rates of the three substrates became apparent.

Aminopeptidases↗
PubMed1983

Aminopeptidases of the rat prostatic complex and seminal vesicles: secretion and effect of castration.

Aminopeptidases hydrolysing L-leucine-beta-naphthylamine (LeuNA), L-alanine-beta-naphthylamide (AlaNA) and L-arginine-beta-naphthylamide (ArgNA) were studied in tissue homogenates of various parts of the rat prostatic complex and seminal vesicles after castration as well as in their 'cellular' and 'secretory' compartments. The 'secretory' fluids were also fractionated by chromatofocusing to reveal the enzymes possibly secreted by the glands. After castration the ventral prostate showed a temporary increase of LeuNA and a continuous rise of AlaNA hydrolysis, while ArgNA hydrolysis remained unaltered. Its 'secretory' fluid contained high hydrolysis rates of the substrates and chromatofocusing revealed two activity peaks with pI values of 6.2 and 5.4. In the lateral prostate no activity changes were recorded after castration. The 'secretory' fluid contained low enzyme activities and the fractionation showed one enzyme activity with pI 5.4. The posterior prostate, coagulating glands and seminal vesicles showed high hydrolysis rates of ArgNA with a rapid decline after castration. The 'secretory' fluid of posterior prostate contained rather low activity levels and in chromatofocusing two activity peaks with pI values of 6.2 and 5.4 were found. The 'secretory' fluid of coagulating gland contained a strong hydrolysis of ArgNA and chromatofocusing gave two activities with pI values of 5.4 and 5.1. The latter seemed to hydrolyse only ArgNA. The viscous secretion of the seminal vesicles contained very low aminopeptidase activities and chromatofocusing also showed a poor result. It is concluded that the various parts of the rat prostatic complex differ in their aminopeptidase composition. Only part of these enzymes appears to be secretory and controlled by testosterone. The major contribution to the seminal plasma aminopeptidase activity seems to derive from the ventral prostate and the coagulating gland.

Aminopeptidases↗
PubMed1983

Acid phosphatases in the mouse testis: activity changes during development.

The histology and acid phosphatase activities of the developing testes in the mouse, from 4 days of age until maturity, were analyzed. The specific enzyme activity with p-nitrophenyl phosphate as substrate increased after 3 weeks of age and then reached a plateau. Cobalt and zinc markedly increased the specific activity after the third week. After fractionation the testicular homogenate revealed four acid phosphatases. Enzyme I maintained a high activity during the first 3 weeks but steadily declined thereafter. Enzyme II, present in all age groups, showed a moderate increase after the fourth week. Enzymes III and IV were low and declined further during the first 3 weeks with a subsequent increase. This occurred concomitantly with the appearance of spermatids and mature sperm cells. Changes in enzyme activities seem to reflect the alterations in cellular composition of the testis during the developmental process. Enzymes III and IV were probably associated with spermatids and sperm cells.

Acid Phosphatase↗
PubMed1983

Characterization of three aminopeptidases purified from human placenta.

Three aminopeptidases purified from the human placenta were characterized and compared with each other. Aminopeptidase II1 preferred L-arginine- and L-lysine-beta-naphthylamides or p-nitroanilides as substrate, with low or negligible hydrolysis of other amino acid derivatives. It was inhibited by L-arginine, L-lysine and L-methionine. This enzyme activity was highly sensitive to heat treatment, N-ethylmaleimide, p-chloromercuribenzoate, puromycin, bestatin, epsilon-amino-n-caproic acid (EACA) and EDTA. After EDTA, this enzyme could be reactivated by Co2+. It is concluded that aminopeptidase II1 is identical with arginine aminopeptidase (EC 3.4.11.6) from other mammalian tissues. Aminopeptidase II2 preferred L-alanine-beta-naphthylamide and p-nitroanilide as substrates. It was also able to hydrolyse L-leucine, L-arginine, L-methionine and L-lysine derivatives but only very weakly L-cystine and Bz-L-cysteine substrates. This enzyme was inhibited by L-arginine, L-alanine, L-lysine and most strongly by L-leucine and L-methionine. It was resistant to bestatin and heat treatment but sensitive to EACA. EDTA caused a marked suppression, which could be prevented by Co2+ and Zn2+. These characteristics are reminiscent of those of alanine aminopeptidase (EC 3.4.11.-) found in other tissues. The third enzyme was the only one clearly particle bound and was therefore called PB-aminopeptidase. It preferred L-leucine derivatives as substrate but also readily hydrolysed other amino acid-beta-naphthylamides and p-nitroanilides including L-cystine and Bz-L-cysteine substrates. Among the amino acids L-cysteine, L-leucine and L-methionine were inhibitory. Bestatin and thiol reagents were without effect and EACA was only moderately inhibitory. EDTA caused a strong suppression, which could be prevented by Co2+ and Zn2+. These properties are equal to those previously described for the placental cystine aminopeptidase (oxytocinase) (EC 3.4.11.3). All three enzymes had an optimum close to neutral pH but apparent differences in their Km and Vmax values with various substrates. These findings suggest that the three purified aminopeptidases are distinct enzymes. Two of these (aminopeptidases II1 and II2) have not previously been isolated and characterized in the human placenta.

Amino Acids↗
PubMed1982

Acid phosphatases of dog testis and a Leydig cell tumor.

Acid phosphatase activities were measured with five different substrates after fractionation with Sepharose 6B and DE-52 cellulose chromatography of homogenate from normal adult dog testis and a testicular tumor. The tumor showed a positive 3 beta-hydroxysteroid dehydrogenase reaction and was diagnosed as a Leydig cell adenoma. The fractionations gave three separate enzyme activities in the normal testis and two enzymes in the tumor. All were sensitive to sodium fluoride, but differed from each other in pH-optima and the response to Co2+ and Zn2+. Enzymes I and II were identical in both tissues. The latter with a smaller molecular weight was activated by Zn2+ but not by Co2+ and had slightly higher pH-optimum (4.5) than enzyme I (optimum at pH 3.5). The third enzyme was activated by Co2+ and Zn2+ and had the highest pH-optimum (pH 5.5). It was called enzyme IV due to its resemblance to a similar activity in other mammalian species.

Acid Phosphatase↗
PubMed1980

Fractionation and characterization of cystine aminopeptidase (oxytocinase) and arylamidase of human serum during pregnancy.

Cystine aminopeptidase and arylamidase activities in human serum were determined by enzymic hydrolysis of L-cystine-di-beta-naphthylamide (CysNA) and L-leucine-beta-naphthylamide (LeuNA), respectively. The activities of both enzymes increased during pregnancy, cystine aminopeptidase 12.5-fold and arylamidase 8.3-fold. Serum CysNA and LeuNA hydrolysing aminopeptidases were separated by gel filtration on Sepharose 6B. Serum from non-pregnant women (control) contained arylamidase (Ic), which hydrolysed LeuNA and (weakly) CysNA, and cystine aminopeptidase II, hydrolysing only CysNA. During pregnancy a new enzyme appeared in maternal serum and showed cystine aminopeptidase and arylamidase activity (Im). Maternal serum Enzyme(s) I had higher pH optima (6.5 with CysNA; 7.5 with LeuNA) and higher molecular weights (309,000) than arylamidase Ic (pH optima at 5.52-5.5 with CysNA and 7.0 with LeuN; mol.wt approximately 130,000). Arylamidase Ic was more sensitive to L-methionine, but more resistant to heat than maternal serum Enzyme(s) I. Both control and maternal serum Enzyme(s) I were inhibited by EDTA, but were re-activated by Zn2+ and Co2+ with LeuNA and CysNA as substrates and by Ni2+ with CysNA. Cystine aminopeptidase Im and arylamidase Im may be a single enzyme although differences were obtained in pH optima and reactivation by Ni2+ after EDTA treatment. Since maternal serum Enzyme(s) I had biochemical characteristics similar to those of placental aminopeptidase(s) I, it is suggested that the activities are of placental origin. Cystine aminopeptidase II appeared in all sera. It differed clearly from both maternal and control serum enzyme(s) I: it had the lowest molecular weight (76,000), a different pH optimum (6.0) and was resistant to EDTA and L-methionine. It was not as effectively inhibited by Ni2+ as was Enzyme(s) I.

Aminopeptidases↗
PubMed1980

Testicular acid phosphatases in cattle, sheep, guinea pig, and rabbit.

Acid phosphatase activities were measured with five different substrates in the total homogenates as well as after gel filtration on Sepharose 6B and cellulose chromatography of bull, guinea pig, rabbit, and ram testes. The response of the hydrolysis rate to NaF (5 mmol/l), Co2+ (5 mmol/l) and Zn2+ (5 mmol/l) was also tested. In the total homogenate the hydrolysis of p-nitrophenyl phosphate was markedly activated by Co2+, while in the presence of Zn2+ an activation was recorded in guinea pig and some inhibition in the bull, rabbit, and ram testes. NaF caused a decline in the total acid phosphatase activity, particularly in guinea pig and ram. The gel filtration resulted in three separate activity peaks with p-NPP and beta-NP as substrates. N-ASBI-P, alpha-NP, and Tym-P gave only two peaks. After subsequent cellulose chromatography of the activities only peak II gave rise to two further activities. Peak I of gel filtration (enzyme I) was able to hydrolyze all substrates tested and was highly sensitive to NaF. Peak I of cellulose chromatography (enzyme II) also hydrolyzed p-NPP and beta-NP. It was rather resistant to NaF but sensitive to Zn2+. It was slightly activated by Co2+. Peak II of cellulose chromatography (enzyme IV) hydrolyzed only p-NPP and was markedly activated by Co2+ and Zn2+. The adult testes of bull, guinea pig, rabbit, and ram have a closely similar testicular acid phosphatase pattern. Due to relative differences in the concentrations of the four enzymes in the tissue, varying activity levels are recorded in the presence of different substrate and modifier combinations.

Acid Phosphatase↗
PubMed1979

Fractionation and characterization of cystine aminopeptidase (oxytocinase) and arylamidase of the human placenta.

Three activity peaks hydrolysing L-cystine-di-beta-naphthylamide (CysNA) and two activities hydrolysing L-leucine-beta-naphthylamide (LeuNA) were separated by gel filtration on Sepharose 6B from human placental tissue. The enzyme activities in the void volume and the solubilized enzyme activities with both substrates apparently are bound and free forms of the same enzymes (I) since detergent treatment caused a total disappearance of the activities in the void volume. The second distinct enzyme (II) was highly soluble and detected only with CysNA. The particle-bound enzyme(s) had a pH optimum at 6.5 with CysNA and at about 7.5 with LeuNA. They were highly sensitive to EDTA, could be reactivated by Co2+ and Zn2+ and were more sensitive to Ni2+ and L-methionine than the soluble enzyme II. The former enzyme(s) tolerated thermal treatment better than the soluble enzyme II. The solubilized free enzyme(s) I had a molecular weight of about 309,000. The soluble enzyme II was resistant to EDTA. Its optimum was at pH 6.0 and an estimate of 76,000 for the molecular weight was obtained.

2,2'-Dipyridyl↗