Proteins of chicken, duck, and turkey egg white

EN,

DUCK,

AND

TURKEY

E(

by • D U C A Y , T. F. S U G I H A 1 2 A AND 12.

esearch Laboratory*, Albany, Calif. (U.5

he c h e m i c a l p r o p e r t i e s a n d biological a~ t a n t to s t u d y t h e c o m p a r a t i v e b i o c h e n c o m p a r i s o n s of v a r i o u s a v i a n egg whi AND H A R T S E L L 2 h a v e r e p o r t e d on t h e bacteriolc [ically. I n b o t th s t u d i e s la Its of different w h i t e s as d e t e r m i n e d bacteriologically. were f o u n d . roe are r e p o r t i n g o n t h e c o m p a r a t i v e c o n t e n t s of l y s o z y m e (be (bacteriolytic 3, ical i r o n - b i n d i n g c a p a c i t y ) s, o v o m u c o i d ( t r y p s i n - i n h i b i t i n g capacity) c ~ecies. T a b l e I., r i b e n z o a t e t i t r a t i o n for s u l f h y d r y l groups)3, 4 of s e v e r a l specie of m a n y c h i c k e n w h i t e s a n d of d u p l i c a t e pooled s a m p l e s of t u r k e y , P e egg white. T h e l y s o z y m e c o n t e n t s are c o m p a r a b l e w i t h tt]h o s e repot ~ELLz T h e r e l a t i v e c o n a l b u m i n c o n t e n t s a r e of t h e s a m e o r d e r as r )ecies were ~cn 1, b u t lower a m o u n t s a n d g r e a t e r differences b e t w e e n sp~ tts of t h e d u c k w h i t e s were h i g h e r t h a n t h o s e of t u r k e y or chicken. ct

proteins gg w h i t e s made by lysozyme s between nalbumin (p-chlorote a v e r a g e d Mallard ELIS AND ~AIN AND

vomucoid

TABLE I D a t a in p e r c e n t a g e , d r y - w e i g h t bbasi asis* Cysteim Bird

Lysozym¢

ConaIbumin

Ovomuvoid Native

Chicken* ** Turkeyt Peking duckt Mallard duckt

3.4 1.84 0.94 0.86

12

ii

0.46

II

II

0.42

2.8

15

o.32

2. 9

15

0.28

E

0.62 0.47 0.37 0.29

• S t a n d a r d s p r e v i o u s l y described s. G u a n i d i n e - H C 1 as p r e v i o u s l y described*. whiter • ** A v e r a g e s f r o m 12 or m o r e different pooled s a m pties (4 t o 8 i n d i v i d u a l eggg whites). A v e r a g e s of d u p l i c a t e pooled s a m p l e s . • *

h e w h i t e s w a s f o u n d in t h e s u l f h y d r y l , or cysteine, c o n t e n t s . A n i m p o r t a n t difference b e t:ween w e e n tthe ound I n c h i c k e n egg white, all t h e cyvsteine e x i s t s in t h e o v a l b u m i n c o m p o n e n t a n d t h e v a l u e s ffouI fo h a v e b e e n s h o w n to be e q u i v a l e nttt to t h r e e s u l f h y d r y l g r o u p s p e r m o l in t h e n a t i v e p r o t e i n a n d four mturation .~ase in s u l f h y d r y l g r o u p s in t h e chicken e g g w h i t e a f t e r denaturati~ in t h e d e n a t u r e d 4. T h e large increase ltion w a s n o t o b s e r v e d in t h e o t h e r w hrites i t e s (Table I). T h e s e r e s u l t s m a y be i n t e r p r e t e d as a demonstrati~ chicke of different m o l e c u l a r s t r u c t u r e inu t h e t u r k e y a n d d u c k o v a l b u m i n s as c o m p a r e d to t h o s e of chicken. O n e p r e p a r a t i o n of semipurified t u r k e y o v a l b u m i n g a v e i d e n t i c a l s u l f h y d r y l t i t r a t i o n s before aaln d a f t e r d e n a t u r a t i o n , t h u s confirmi~n g t h e r e s u l t s f o u n d o n t h e t u r k e y white. Also, f r o m t h e o v a l b u m i n c o n t e n t s r e p o r t e d b y BAIN AND DEUTSCHx a n d on t h e a s s u m p t i o n t h a t t h e o v a l b u m i n s of t h e o t h e r species h a v e m o l e c u l a r w e i g h t s similar to t h e c h i c k e n s ' (46,ooo ± i o % ) , one c a n calculate t h r e e r k e y o v a l b u m i n a n d t w o p e r m o l for d u c k o v a l b u m i n . s u l f h y d r y l g r o u p s p e r tool for turke~ o S t u d i e s of t h i s n a t u r e m u s t be c a r e f u l l y a p p r a i s e d w i t h r e g a r d to t h e m e t h o d s a n d a s s u m p t iions tre in t e r m s of t h e p r o t e i n s of c h i c k e n w h i t e as s t a n d a r d s a n d lar ge e m p l o y e d . T h e r e p o r t e d r e s u l t s are a c t i v i t i e s m a y exist. F u r t h e r s t u d i e s o n t h e c o m p~arative arati' differences in m o l e c u l a r w e i g h t s a~nd n d specific sr w a n t i - h e m a g g l u t i n i n activities a n d] t h e c u l i n a r y a n d f o a m i n g p r o p e r t i e s of t h e v a r i o u s egg w h i t e s will be r e p o r t e d elsewhere. U " B u r e a u of A g r i c u l t u r a l a n d I n d u s t r i a l C h e m i s t r y , A g r i c u l t u r a l R e s e a r c h A d m i n i s t r a t i o n , U.S. D e p a r t m e n t of A g r i c u l t u r e .

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Biol. Chem., i 7 i (1947) 53 I. L, Prec. Soc. Exptl. Biol. Med., 76 (195 SILVA AND L. R. M A C D O N N E L L , Poult~ D R . E . F E E N E Y , Arch. Biochem. Biopi Recei

52) 639. ) 288. r 2 t h , 1953

IN T H E R A T OF G U A N Y L I kBELLED W YVIJ[T I H .L-1-L ssp* --.L-

..L,.L~.IJJL~.IL~JL~.L~JLJ

by P A U L M. R O L L , H E R B E R T

WEINFELD

AND G E O R G E

BOSW~

earch, N e w Y~ ; Laboratories o/the Sloan-Kettering Institute/or Cancer Reseat,

WN

U.S.A .)

arsors b u t t h a s b e e n e s t a b l i s h e d t h a t , in t h e rat, g u a n i n e a n d guano,, u a n o s i n e a r e ve :uauylic a c i d s s e r v e as effective p r e c u r s o r s of p e n t o s e nuclq :leic acid (1~ ample, no ,oration of g u a n i n e 1, ~ or g u a n o s i n e s, labelled w i t h 16N, into t h e P N A gl combined of t h e v i s c e r a of t h e r a t c o u l d be d e t e c t e d , a l t h o u g h ssub~ u b s e q u e n t in gith these rends, labelled w i t h t h e m o r e s e n s i t i v e t r a c e r 14C, r e v e a l e d t h a t each r a t e d to a extent4, 5. I n c o n t r a s t , it w a s f o u n d t h a t t h e P N A g u a n i n e of q the rat ~tensively a n d " b ''6 d f r o m y e a s t g u a n y l i c acid, w h i c h w a s p r e s u m a b l y a m i x t umr e of guam. W da. ~s, s ~ lh J .o UW L.LL~ O l~U.l.ll~.mL 'w a s n t l y c o n s i d e r e d t o be t h e 2" a n d 3'-isomers) a n d l a t e r it w wa: w/ Jn. tL.IJ.~L t ;omer w m o r p o r a t e d 7. I t m u s t be n o t e d t h a t in all of t h e a b o v e sstudi, t u d i e s o n l y t h e g u a n i n e m o i e t y of t h e Listered n u c l e o s i d e s a n d n u cflee l e o t i d e s h a d b e e n labelled so t h a t o n l y t h i s p o r t i o n of t h e m molect olecule w a s t rraced. a c e d . H o w e v e r , t h e r e s u l t s of t h e e x p e r i m e n t s do d e m o nms t r a t e t h a t g u a n i n e in nnucleotide ucleoti l i n k a g e is m u c h m o r e e x t e n s i v e l y utilized t h a n is t h e p u r i n e , aatn d it m a y be c o n c l u d e d t h a t t h e m e c h am i s m for t h e i n c o r p o r a t i o n of g u a n y l i c a c i d s does n o t i n vwo l v e a s i m p l e liberation of eit~:her or t h e free p u r i n e p r i o r t o t h e i n c o r p o r a t i o n . A p)ossible m e c h a n i s m w o u l d i n v o l v e t h e n ulcleoside c t h e incor ~orporation of t h e i n t a c t n u c l e o t i d e s d u r i n g t h e b i o s y n t h e s i s of o t h e P N A . T o t e s t t h i s possibilit )ossibihty, g u a n y llic i c a c i d s " a " a n d " b " were p r e p a r e d w h i c h were labelled umn i f o r m l y w i t h 15N, u n i f o r m l y, wi with 14C a n ~d d w i t h s a p a n d were a d m i n i s t e r e d to r a t s . T h e r e s u l t s of t h e u t i l i z a t i o n of t h e p h o s pJhho r u s of t h e s e n u c l e o t i d e s are p r e s e n t e d here. T?he h e p r e p a r a t i o n of t h e m u l t i p l y labelled g u a n y l i c acids w a s carried c o u t b y isolating t h e n u cd e otides b y ion e x c h a n g e o n D o w e x - i s ( N a t i o n a l A l u m i n a t e Corpo~ )oration, U.S.A.) f r o m h y d r o l yrsates sat 5 P labelled nucleic of nucleic :leic a c i d s labelled w i t h 14 C * * 15 N 5 a n d 8~ P. T h e 5asp n acid w a s isolated b y • ssodiu odium chloride e x t r a c t i o n f r o m y e a s t w h i c h h a d b e e n g r o w n in a m e d i u m c o n t a i n i n g 3~p*** a n d w a s purified b y p r e c i p i t a t i o n f r o m a qLueous s o l u t i o n w i t h glacial acetic acid. T h e t h r e e s e p a r a t e l yr labell labelled s a m p l e s of e a c h n u e l e o t i d e were c o m b i n e d a n d were a d m i n i s t e r e d to m a l e S h e r m a n s t r a i n r a t s of 2 5 0 - 3 o o g b y i n t r a p e r i t o n e a l injqection. E a c h r a t received o. 4 m M p e r k i l o g r a m of b o d y w e i g h t in t h r e e e q u a l doses a t a b o u t t w o - h o~ur u r iinter~ n t e r vwa l s , a n d t h e a n i m a l s were sacrificed a p p r o x i m a t e l[y y twent yf o u r h o u r s a f t e r t h e first injection.i. T h e spleen, liver, k i d n e y s , t e s t e s a n d s m a l l i n t e s t i n e were r e m o v e d a n d were frozen o n dry-ice. T h Le e t i s s u e s were c o m b i n e d a n d were h o m o g e n i z e d in ice-cold lIO O% t r i e h l o r o a c e t i c acid (TCA). T h e r•eesidue s i d u e w a s d e h y d r a t e d w i t h alcohol a n d e t h e r to yield a d r y tiss tissue p o w d e r f r o m w h i c h r i b o n u c l e o t i d]es e s were o b t a i n e d b y a modification of t h e SCHMIDT-THANNHAUSI;ER p r o c e d u r e 1° a n d were s e p a r a t e d b) 3y ion e x c h a n g e on D o w e x - i . S a m p l e s of t h e s o l u t i o n s of t h e n u c l e o t i d e s were p l a t e d on a l u m i n u m p l a n c h e t s a n d t h e aactivi ctivity d u e to sap w a s d e t e r m i n e d in a n int n t e r n a l Geiger-Mfiller flow c o u n t e r ( R a d i a t i o n C o u n t e r Laboratoric )oratories, m a r k 12, m o d e l I, helium-isobutane a n e gas), w i t h t h e u s e of a n a l u m i n u m shield w h i c h h a d b e e n foul found * T h i s i n v e s t i g a t i o n w a s s u p)p o r t e d b y g r a n t s f r o m t h e N a t i o n a l C a n c e r I n s t i t u t e , N a t i o rn a l I n s t i t u t e s of H e a l t h , U n i t e d S t a~tes t e s P u b l i c H e a l t h Service, f r o m t h e A t o m i c E n e r g y C o m m i s s i co n , C o n t r a c t No. A T ( 3 o - i ) - 9 i o , a n d f r o m t h e N u t r i t i o n F o u n d a t i o n , Inc. ** O b t a i n e d f r o m S c h w a r z L a bboratories, o r a t oories r i e Inc., N e w York, U.S.A. *** O b t a i n e d f r o m O a k R i d g e N~lational a t i o n a l L a b o r a t o r y , T e n n e s s e e , U.S.A., as i n o r g a n i c p h o s p h a t e .