Protein phosphokinase reactions in mammalian testis. Stimulatory effects of adenosine 3′:5′-cyclic monophosphate on the phosphorylation of basic proteins

Reddi, A. Hari; Ewing, Larry L.; Williams-Ashman, H.G.

doi:10.1042/bj1220333

Cited by 50 publications

(13 citation statements)

References 34 publications

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“…Assay of protein kinase was performed by a minor modification of the method of Reddit et al [8], using histone F2b as a substrate in the presence or absence of 4 I.~M cyclic AMP.…”

Section: Introductionmentioning

confidence: 99%

Adenosine 3′,5′‐monophosphate‐dependent protein kinase of leydig cells: In vitro activation and relationship to gonadotropin action upon cyclic AMP and steroidogenesis

1976

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“…Assay of protein kinase was performed by a minor modification of the method of Reddit et al [8], using histone F2b as a substrate in the presence or absence of 4 I.~M cyclic AMP.…”

Section: Introductionmentioning

confidence: 99%

Adenosine 3′,5′‐monophosphate‐dependent protein kinase of leydig cells: In vitro activation and relationship to gonadotropin action upon cyclic AMP and steroidogenesis

1976

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“…Reddi et al (1971) have shown that the soluble fraction of whole testis homogenates contained the protein kinase enzyme and that cyclic AMP markedly stimulated the enzyme with histone as substrate. Bernard & Wasserman (1974) also reported cyclic AMP-dependent protein kinases in rat testis interstitial tissue and seminiferous tubules.…”

Section: Discussionmentioning

confidence: 99%

“…The effect of LH -on protein iasesI in the estis has not been-investigated previonsly4 but!erlier ,work on'the testis has shown that cyclic AMPdependent protein kinases are presnt (Reddi et al,, 1971) and that FSH can stimulate protein 1kinawscti-vity in the-seminiferous tubules (Means-at (4., 1974). …”

Section: Vol 154mentioning

confidence: 99%

Protein kinase activity in rat testis interstitial tissue. Effect of luteinizing hormone and other factors

Cooke¹,

Kemp²

1976

Biochemical Journal

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Protein kinase:activity was determined in subcellular fractions of-rat testis interstitial tissue after-incubation ofthe-intact tissue with LH (luteinizing ¶hormone) in vitro. Various factors that might have changed the activity of this .enzyme during preparation of the fractions beforeassay were also investigated. The following results were obtained. 1.LLH and 34sobutyl-b 1methylxanthine (a phosphodiesterase inhibitor) added together-during incubation of the interstitial tissue,caused a twofold increasein the protein kinase-activity in :the total tissue homogenate and subcellular fractions (12000gx5min pellet and 0 5O0fgx60min supernatant and pellet). 2. A decrease of approx. 40% in -the total amount of proteinkinaserecovered in the solublefraction (1O5Q00gsupernatant}occurred in tissue incubated with -LH and 3-isobutyl-1-methylxanthine when compared with the controls. No change in total activity was found in the other fractions. 3. LH Sand 3-isobutyl4ln-methylxanthine caused;an increase in cyclic AMP concentration in the soluble fraction (from, 30±6 to 450±+4prnio1/mg of protein, -means±S.E.M., -n=4), but there -waslittle orno inceaseinthe paitiqulate.fractions [from 9± to 13 + &pmol/mg ofprotein (n =3) and -from,6±2)to 23+1l1pmol/mg of protein,(n =-3) in thle 12000gand-.105000g -pellets respectively]. 4. Addition of 3-isobutyl.l-methylxanthine alone had little effect -on protein kinase activity or cyclic-AMP concentrations. 5. Little or no protein kinase .activity could be-demonstrated in subcellular particulate fractions-unless Triton X-I00 was added; the effect of this detergent was shown.to;be at least partly due to the inhibition -of adenosine triphosphatase activity. 6. In the presence of Triton X-100 approx. 57% of the total protein Ekinase activity in the homogenate was found in the 105 OOOg supernatant compared with 11 %in-the 1-OSOOg pellet and 32% in the 4200g-pellet. 7. In contrast mith adipose-tissue protein kinase [Corbin et al. (1973)-J. Biol. Chem. 248, 1813-1821 the.relative amounts -of cyclic AMP-independent and -dependent enzyme -were -not affected by dilution of the interstitial-tissue fractions. NaCl (0.5M) decreased the estimated total amount of protein kinase activity.

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“…This gives a calculated minimal molecular weight of 17,982, a value in close agreement with a value of around 17,900 computed (4) from the results of physical measurements. It is noteworthy that the basic protein of guinea pig seminal vesicle secretion is an excellent substrate for cyclic AMP-stimulable protein phosphokinases (14). Nevertheless, the protein as isolated is entirely devoid of phosphorus; ultraviolet spectra and analyses for bound carbohydrates suggested that the basic protein is devoid of prosthetic groups.…”

Section: Methodsmentioning

confidence: 99%

“…The purified basic protein migrated towards the anode at pH 9.3 or greater, and exhibited only a single band on electrophoresis in either direction at different pH values from pH 6-10. A more thorough examination of the amino-acid composition of the native basic protein (14), involving slight modification of the procedures used earlier (4), indicated that the amino-acid composition of this protein is: Asp,,, Thr6, Sern, Glu28, Gly19, Ala6, Val13, Met6, 11e2, LeuM4, Phe6, Lys16, His2, Argii, Try,, with 21 amide-NH2 residues and the complete absence of Pro, Tyr, and cyst(e)ine. This gives a calculated minimal molecular weight of 17,982, a value in close agreement with a value of around 17,900 computed (4) from the results of physical measurements.…”

Section: Methodsmentioning

confidence: 99%

Transamidase Reactions Involved in the Enzymic Coagulation of Semen: Isolation of γ-Glutamyl-ε-Lysine Dipeptide from Clotted Secretion Protein of Guinea Pig Seminal Vesicle

Williams-Ashman

Notides

Pabalan

et al. 1972

Proc. Natl. Acad. Sci. U.S.A.

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New supportive evidence is advanced in favor of the hypothesis that the enzymic coagulation of guinea pig semen involves transamidase reactions that result in the formation of 7y-glutamyl-e-lysine intermolecular cross linkages between molecules of a basic protein in seminal vesicle secretion. The dipeptide -y-glutamyl-elysine was isolated in large quantities from proteolytic digests of the coagulated basic vesicular secretion protein that comprises the seminal clot.The seminal vesicle secretions of guinea pigs (1-4), rats (4-9), and mice (10) contain a small group of bulk proteins that are easily separable from one another by chromatographic and electrophoretic procedures. One of these proteins is strongly basic, and can be coagulated by an enzyme from the anterior prostate (coagulating) gland, but not by thrombin (4, 5). This enzymic coagulation process is centrally involved in the postejaculatory clotting of rodent seminal plasma that results in the formation of a vaginal plug when semen is catapulted into the female genital tract at coitus (1,4,5).In 1967, Notides and Williams-Ashman (4) purified the basic coagulable protein from guinea pig vesicular secretion to a state of homogeneity. They observed that, after hydrolysis in strong acid, the amino-acid composition of this basic protein [which is devoid of proline, tyrosine, and cyst(e)ine] was remarkably similar to the amino-acid composition of washed coagulated material formed by the action of the anterior prostate enzyme on a crude mixture of the saline-soluble bulk proteins of seminal vesicle secretion. The native vesicular secretion basic protein has a molecular weight of roughly 17,900, whereas the molecular weight of the coagulated protein is not known. It was observed (4) that clotting of the vesicular secretion basic protein by the anterior prostate enzyme reduced the amide-NH2 from a value of about 21 residues per 17,900 molecular weight to a value of roughly 12 amide-NH2 residues (the number of amide residues represented by glutaminyl or asparaginyl groups was not determined). The enzymically clotted basic protein is not soluble in 8 M urea or in 6 M guanidinium chloride in the absence or presence of thiol compounds at neutral pH (see ref. 1). These facts, taken together, led Notides and Williams-Ashman (4) to postulate that the enzymic clotting of the basic seminal vesicle secretion protein involved the formation of intermolecular linkages similar to those formed in the fibrinoligase (thrombin-activated fibrin stabilizing factor or activated Factor XIII) reaction of mammalian blood clotting (11-13). It is now well established that fibrinoligase is a transamidating enzyme, which catalyzes the production of selective -y-glutamyl-e-lysine crosslinks between fibrin molecules.Some additional new evidence in support of the hypothesis that the guinea pig seminal clot is formed by an enzymic transamidation reaction is outlined below. Unequivocal proof of the formation of peptide cross linkages is provided by the isolation of 'y-glutamyl-e-lysine d...

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Protein phosphokinase reactions in mammalian testis. Stimulatory effects of adenosine 3′:5′-cyclic monophosphate on the phosphorylation of basic proteins

Cited by 50 publications

References 34 publications

Adenosine 3′,5′‐monophosphate‐dependent protein kinase of leydig cells: In vitro activation and relationship to gonadotropin action upon cyclic AMP and steroidogenesis

Adenosine 3′,5′‐monophosphate‐dependent protein kinase of leydig cells: In vitro activation and relationship to gonadotropin action upon cyclic AMP and steroidogenesis

Protein kinase activity in rat testis interstitial tissue. Effect of luteinizing hormone and other factors

Transamidase Reactions Involved in the Enzymic Coagulation of Semen: Isolation of γ-Glutamyl-ε-Lysine Dipeptide from Clotted Secretion Protein of Guinea Pig Seminal Vesicle

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