1. &#945;-Aminoadipic acid and lysine are increased in the urine of thyroidectomized dog. 2. Pipecolic acid is increased in the urine of rat treated with thyroid extract.
3. Relation between thyroid function and lysine metabolism is discussed.</p

Doi, Genziro

Iwado, Yoshiyuki

Kurahashi, Kengo

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 12, Issue 4 1958 Article 1DECEMBER 1958Studies on thyroid func-tion &#x2161;. Effectof thyroidectomy and thyroid extract on theurinary amino acids of animalsKengo Kurahashi∗ Yoshiyuki Iwado†Genziro Doi‡∗Okayama University,†Okayama University,‡Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Studies on thyroid func-tion &#x2161;. Effectof thyroidectomy and thyroid extract on theurinary amino acids of animals∗Kengo Kurahashi, Yoshiyuki Iwado, and Genziro DoiAbstract1. α-Aminoadipic acid and lysine are increased in the urine of thyroidectomized dog. 2.Pipecolic acid is increased in the urine of rat treated with thyroid extract. 3. Relation betweenthyroid function and lysine metabolism is discued.∗Copyright c©OKAYAMA UNIVERSITY MEDICAL SCHOOLActa Med. Okayama 12, 287-292 (1958)STUDIES ON THYROID FUNCTION*11. EFFECT OF THYROIDECTOMY AND THYROID EXTRACTON THE URINARY AMINO ACIDS OF ANIMALSKengo KURAHASHI, Yoshiyuki IWADO and Genziro DOlDepartment of Biochemistry, Okayama University Medical SchoolOkayama, Japan (Director: Prof. S. Mizuhara)Received for publication, August 11, 1958In the previous papersl , 2. 3.4, it has been reported that ,a-alanine, sar-cosine, lysine and its metabolic intermediates are increased in the urineof both hyper- and hypothyroid patients, and the administration of thy-roid extracts to the rabbits has led to the similar changes in the aminoacid pattern of the urine and the muscle nitrogen pool. In this communi-cation, further studies on the relation between lysine metabolism andthyroid function were carried out by using thyroidectomized and thy-roxine treated animals.METHODSThyroidectomy was carried out by the procedure stated in BOMSKOV'shandbook 5. For the induction of hyperthyroid state, 0.25 ml per day of"Thyradin" (Teikoku Zoki CO., Ltd.) was injected intraperitoneally intorats for seven days. One ml of Thyradin corresponds to 0.2 gm of freshthyroid. Urine was collected under toluene and desalted by the methodof CARSTEN 6• Ion exchangers used were "Daiaion" SK-l and SA-lOO(Mitsubishi Kasei CO., Ltd.). The dipole ion fraction thus obtained waschromatographed in two ways on paper (Toyoroshi No. 50). Solvents usedthroughout this series of study were n-butanol-acetic acid-water (5:1.2: 5 VIV) and phenol-water (4: 1; 0.3 % NH~ in the chamber).0.1 % ninhydrin in water-saturated n-butanol was used for color develop-ment.Since most of the amino acids ever appeared on paper have alreadybeen identified in the previous reportsl - 4, further identification was un-necessary except one amino acid in this experiment.* This work was supported by a research grant from the Ministry of Education.2871Kurahashi et al.: Studies on thyroid func-tion &#8545;. Effect of thyroidectomy andProduced by The Berkeley Electronic Press, 1958288 K. KURAHASHI, Y.IwADo and G. DOlRESULTS(I) Effect of thyroidectomy on the urinary amino acid pattern of ayoung dog:Fig. 1 shows the paper chromatogram of urinary amino acids of anormal puppy weighing 3.1 kg. The little dog was fed on cow milk.r-Aminobutyric acid (spot e) was the first amino acid met in the animalurine ever studied in this laboratory. Spot c appeared between j9-alanine8 7 6 S 4 3 2. /BUTANOL-ACETIC ~(__1. Alanine2. Arginine3. Aspartic acid5. Glutamic acid6. Glycine7. Histidine8. Hydroxyproline9. Isoleucine10. Leucine13. Phenylalanine14. Proline15. Serine16. Threonine18. Tyrosine19. Valinea. a-Aminoadipic acidb. ,B-Alaninec. Unknownd. ,B·Aminoisobutyric acide. ,-Aminobutyric acidf. Sarcosineg. Il.Aminovaleric acidFig. 1. Urinary Amino Acids of Normal Puppy Fed Cow Milkand hydroxyproline was the only amino acid yet unidentified. The rela-tive situation between hydroxyproline and spot c on paper is similar tothat of proline and pipecolic acid. So spot c was considered to be hy-droxypipecolic acid, and expectedly it resisted the nitrite treatment asshown in Fig. 2. This spot c, however has no relation with the thyroidfunction, so further identification was not carried out with the knownsample.Fig. 3 shows the urinary amino acid pattern of the dog forty daysafter thyroidectomy. Compared with Fig. 1, the remarkable changes ofthe chromatog~amare the increase of a-aminoadipic acid (spot a) and the2Acta Medica Okayama, Vol. 12 [1958], Iss. 4, Art. 1http://escholarship.lib.okayama-u.ac.jp/amo/vol12/iss4/1Studies on Thyroid Function, -.. 0.8, , f'tIt.-e~ 8. Hydroxyproline0.7 c. Unknownf. Sarcosine0.6r-t0T~f0.2 0.1+--Butanol-AceticFig. 2. Urinary Amino Acids of Normal DogAfter Nitrite TreatmentFig. 3. Urinary Amino Acids of Thyoidectomized Dog2893Kurahashi et al.: Studies on thyroid func-tion &#8545;. Effect of thyroidectomy andProduced by The Berkeley Electronic Press, 1958290 K. KURAHASHI, Y. IWADO and G. DOlappearance of lysine (spot 11), thus indicating the intimate relation be-tween thyroid function and lysine metabolism..(lI) Effect of administration of thyroid extract on the urinaryamino acid pattern of rats:Fig. 4 shows the paper chromatogram of urinary amino acids of anormal rat, and Fig. 5 shows that of the same rat after Thyradin treat-ment. A large spot of pipecolic acid has appeared after Thyradin treatment. Although pipecolic acid is often contained in the normal rat urine,it appeared only after Thyradin treatment in this case.DISCUSSIONIt is well known that pipecolic acid and a -aminoadipic acid are themetabolites of lysine in the animal body 7.J~o °14 0·9 1- Alanine0 19dOOO ~ll 3. Aspartic Acid0.8 5. Glutamic Acid6. GlycineC CS Q 7 0.7. 7. Histidine@a '.\ 8. Hydroxyproline9. Isoleucine@ " 10. Leucineo 1 \ \ 0.6 11. Lysine16~ , 14. Proline0.5 15. Serine\:)6 16. Threonine0.4 19. ValinelsO a. ft·Alanineb. Sarcosine,.. , 0.3 c. I'.Aminobutyric Acidf O@ :0; d. {i.Aminovaleric Acid'0 ,~ ...., \ 0.2 f. a-Aminoadipic Acid5 ' ... I, ,b,~,11,, \3 , 00.6 0.5 0.4 0.3 0.2 0.1«Butanol-AceticFig. 4. Urinary Amino Acids of Normal Rat4Acta Medica Okayama, Vol. 12 [1958], Iss. 4, Art. 1http://escholarship.lib.okayama-u.ac.jp/amo/vol12/iss4/1Studies on Thyroid Function 291r\16\Q}60.90.80.70.60.50.41. Alanine3. Aspartic Acid5. Glutamic Acid6. Glycine14. Proline16. Threoninea. ,a-Alaninec. ,/,-Aminobutyric Acidd. o-Aminovaleric Acide. Pipecolic Acid€?l::::? 0.350.23'=:> h0Butanol-AceticFig. 5. Urinary Amino Acids of Thyradin Treated Rat0.6 0.5 0.4 0.3 0.2 0.1CThe present experiment and others1,-4 showed that lysine and its me-tabolic intermediates were increased in the urine of hyper- and hypothy-roid animals.These facts may suggest that thyroxine is a control agent for theutilization of lysine in the animal body.This assumption is in accord with the findings in that the cyto-chrome c contents of thyroidectomized rats is less than one half of thenormal 8, and the lysine content of this important electron carrier is sur-prisingly high 9.Although spot c (Figs. 1-3) is assumed to be hydroxypipecolic acidwhich is derived from hydroxylysine, it is not clear whether its origincomes from cow milk or from tissue constituent. This is the first report,and at any rat, it indicates the occurrence of hydroxypipecolic acid in theanimal urine.5Kurahashi et al.: Studies on thyroid func-tion &#8545;. Effect of thyroidectomy andProduced by The Berkeley Electronic Press, 1958292 K. KURAHASHI, Y.lwADo and G. DOlSUMMARY1. a-Aminoadipic acid and lysine are increased in the urine of thyroi"dectomized dog.2. Pipecolic acid is increased in the urine of rat treated with thyroidextract.3. Relation between thyroid function and lysine metabolism is discus-sed.REFERENCES1. SONODA. Y. ]. lap. Biochem. Soc. 29, 80, 19572. SONODA, Y. Proc. Japan Academy 33, 162, 19573. SONODA, Y. ]. lap. Biochem. Soc. 29, 83, 19574. NAKAMURA, R. : and Negi, Y., Proc. Japan Academy 33, 166, 19575. BOMsKov, C. : Methodik der Hormonforschung, Leipzig, Band I, 149, 19376. CARSTEN, M. E. : ]. Am. Chem. Soc. 74, 5954, 19527. ROTHSTEIN, M. : and Miller, L. 1. : ]. Am. Chem. Soc. 75, 4731, 1953; ibid.,76, 1459, 1954 ; J. BioI. Chem. 206, 243, 19548. DRABKINS, D. 1. : ]. BioI. Chem. 182, 335, 19509. LEAF, G., GILLIES, N. E., and PIRRIE, R. : Biochem. ]. 69, 605, 19586Acta Medica Okayama, Vol. 12 [1958], Iss. 4, Art. 1http://escholarship.lib.okayama-u.ac.jp/amo/vol12/iss4/1

Studies on thyroid function Ⅱ. Effect of thyroidectomy and thyroid extract on the urinary amino acids of animals

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Studies on thyroid func­tion &#8545;. Effect of thyroidectomy and thyroid extract on the urinary amino acids of animals

Abstract

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Okayama University Scientific Achievement Repository

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Studies on thyroid function Ⅱ. Effect of thyroidectomy and thyroid extract on the urinary amino acids of animals