16
J. T. DOLUISIO AND A. S . ~ I A R T I Y
1701.ti
L-Citrullyl-L-prolyl-~~-prolylglycyl-L-phenylalanyl-~-seryl-~acetic acid) ) vas homogeneous on paper chromatogrmiis tixiprolyl-L-phenylalanyl-L-arginine(1-Citrulline Bradykinin) (XX). tariol-acetic acid-water 4 : l :5 , IZr 0.30) and by paper c4ectro-To a solution of the benzyloxycarbonyl nonapeptide methyl phoresis (pyridinc acetate buffer pH 4.0 and ammonium acct:Ltcl ester (XIX) (130 mg.) in methanol (1 nil.) 2 A' sodium hydroxide buffer pH 5.:3) when developed with ninhydrin, Ehrlicli (1'(0.1 ml.) \vas added. Aft'er 1 hr. a t room temperature water (2 dimeth~~lamiiiol~crizalcieh~de) and Sakaguchi reagents. nil.) was added and after another 15 min. the solutio11 was acidi:lnzino .2cid .inaiUsis.--C:ly:Ser:Pro:Phc: Arg: ('it -( h -1 : fied with .V hydrochloric acid and kept, in a refrigerator overnight. 0.8:3 : 2 : 1.1:I .I). The supernatant was decanted and the semi-solid precipitate x i s dried in ZJUCUO over sodium hydroxide. The solid residue thus Acknowledgment.-The author wishes to express hi:, ohtnined (88 mg.) was dissolved in a mixture of acetic acid-nater t u Dr. J1. Bodanszky for the encouragement gratitude ( 5 : l ) (13 ml.) and hydrogenated a t atmospheric pressure for 18 and advice with vhich he supported this work and t o hr. in the presence of 5% palladium on tiariurn sulfate.'* After removal of the catalyst tlie solution n'as freeze-driod. The Dr. J. Fried for his help in the preparation of the manuresidue n-as chromatographed on carhoxyineth~lcellulose, using script. He also is grateful to Mr. J . Alicino and his :I gradually increasing concentration of ammonium acetate for group for the microanalys-es,Mr. 0. I of fl arid p(F-TA1)for tetr~ncyvc~li~~c~~lI('l niitl cupric ions arc given in Table, I and I I I'rom thc foi*mationcur\ c', I'ig. 2 , it -('om- that t h o T alii+ of iz arc' approaching :I \-:tIiw of 2 aiid. I I P I ~ ( Y ~ . t h e \roirhiomctriv ratio of thc c ~ ~ m p l i:,c s2 tc.t i w r l i~i i i ~ molecules pcr cupric. ioii. .lpprosimatct val~ic>> of t IIP qtability coli-tants also may 1~ dcrivcd from thii tormation ciii'vc 11 I
