amino-acid sequence of parvalbumin from rabbit skeletal muscle

Amino-Acid Sequence of Parvalbumin from Rabbit Skeletal MuscleAuthor(s): David L. Enfield, Lowell H. Ericsson, Hubert E. Blum, Edmond H. Fischer and HansNeurathSource: Proceedings of the National Academy of Sciences of the United States of America,Vol. 72, No. 4 (Apr., 1975), pp. 1309-1313Published by: National Academy of SciencesStable URL: http://www.jstor.org/stable/64840 .

Accessed: 03/05/2014 11:03

Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at .http://www.jstor.org/page/info/about/policies/terms.jsp

.JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range ofcontent in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new formsof scholarship. For more information about JSTOR, please contact [email protected].

.

National Academy of Sciences is collaborating with JSTOR to digitize, preserve and extend access toProceedings of the National Academy of Sciences of the United States of America.

http://www.jstor.org

This content downloaded from 130.132.123.28 on Sat, 3 May 2014 11:03:54 AMAll use subject to JSTOR Terms and Conditions

http://www.jstor.org/action/showPublisher?publisherCode=nas

http://www.jstor.org/stable/64840?origin=JSTOR-pdf

http://www.jstor.org/page/info/about/policies/terms.jsp


Proc. Nat. Acad. Sci. USA Vol. 72, No. 4, pp. 1309-1313, April 1975

Amino-Acid Sequence of Parvalbumin fro (calcium-binding protein)

DAVID L. ENFIELD, LOWELL H. ERICSSON, HUBERT AND HANS NEURATH

Department of Biochemistry, University of Washington, Seattle, Wash.

Contributed by Edmond H. Fischer, January 8, 1975

ABSTRACT Determination of the complete amino- acid sequence of rabbit skeletal muscle parvalbumin is described. The sequence of 86 of the 109 total residues was determined automatically by sequenator analyses of peptides obtained after cleavage with CNBr or with trypsin. The positions of the remaining 23 residues were determined by subtractive Edman degradation of t'ryptic and chymotryptic peptides. The protein has an acetylated amino terminus. Comparison of the rabbit parvalbumin with those from carp, hake, and pike and with the calcium-binding subunit of rabbit muscle troponin indicates that these proteins are homologous. Among the parvalbumins a high degree of identity is observed, especially of residues involved in the binding of calcium or in the formation of the hydrophobic core.

The parvalbumins are low molecular weight, acidic, water- soluble, calcium-binding proteins long believed to occur ex- clusively in the white muscle of lower vertebrates and for which no physiological function has been determined as yet (1-11). Amino-acid sequences indicate that they are homolo-

gous (10, 12, 13) and that this homology extends to other muscle calcium-binding proteins, such as troponin C and one of the light chains of myosin (14). The tertiary structure of carp parvalbumin III has been elucidated (15, 16).

Recently, two independent reports have described the isolation of parvalbumins from the skeletal muscle of higher vertebrates, including turtle, chicken, rabbit, and man (17, 18). The retention of these proteins throughout vertebrate evolution suggests that they might be of basic physiological importance. As a first step toward defining their structural and functional relationship to each other and to other muscle proteins, the amino-acid sequence of rabbit parvalbumin is described.

METHODS

Parvalbumin was isolated from rabbit skeletal muscle (17) by Dr. Pavel Lehky. The protein was fragmented and analyzed as shown diagrammatically in Fig. 1. Cleavages with CNBr and with L-l-tosylamido-2-phenylethyl chloromethyl ketone- trypsin were performed as described (10, 19). The CNBr fragments were fractionated by gel filtration (Fig. 2). Enzy- matic digests were subjected to ion-exchange chromatography (Spinco AA-15, Bio-Rad AG1-X2, and AG50W-X2 resins), with gradients of pyridine-acetate buffers. Specific cleavage

Abbreviations: CB, cyanogen bromide fragment; PhS-hydantoin, phenylthiohydantoin; Tp, tryptic peptide; C, chymotryptic peptide; TN-C, the calcium-binding protein of the myofibrillar troponin complex. * Present address: University of Freiburg, Department of Medi- cine, Freiburg im Breisgau, West Germany.

13'

m Rabbit Skeletal Muscle

E. BLUM*, EDMOND H. FISCHER,

98195

adjacent to the arginyl residue was accomplished after succinylation of the e-amino groups (20). The digest was fractionated on Sephadex G-50 "superfine".

Automatic sequence analysis was performed with the Beck- man Sequencer (model 890B), and the silylated phenylthiohydantoin (PhS-hydantoin)-amino acids were identified by gas-liquid chromatography (21). The sequence of the remain-

ing residues was determined by subtractive Edman degradation (22) of tryptic and chymotryptic peptides.

The amino-terminal acetyl group was identified by the procedure of Schmer and Kreil (23). Amino-acid analyses were performed on Beckman (model 120C) or Durrum (model D-500) amino-acid analyzers. Homoserine lactone was con- verted to homoserine before analysis (24). Residues containing amide sidechains were identified by gas-liquid chromatography (21), by high voltage electrophoresis at pH 6.5, or by amino-acid analysis after digestion with aminopeptidase M

(25).

RESULTS

As observed for most other parvalbumins (10, 12, 13), the amino terminus of the rabbit protein is blocked. The entire

sequence was, therefore, derived by analysis of peptides produced by specific cleavages of the polypeptide chain. The amino-acid compositions of the protein and the four major CNBr (CB) fragments are given in Table 1. Digestion of the intact protein with trypsin yielded 17 major peptides. Their amino-acid compositions and positions in the molecule are listed in Table 2. For clarity in presentation of the proof of the structure, residues will be numbered according to the complete amino-acid sequence given in Fig. 3.

Isolation and Sequenator Analyses of CNBr Fragments. The mixture of CNBr fragments was fractionated into five

fractions, CB I to CB V (Fig. 2). B3y sequence analysis all were homogeneous, except CB V, which yielded no sequence. Extended sequenator analyses were carried out on CB II

through CB IV (Fig. 1). The amino-terminal sequence of CB II begins with Val33-Gly-Leu and was determined through Phe7o, except for residues 55, 60-62, and 68, which were identified independently (see below).

Sequenator analysis of CB III yielded 16 residues beginning with Thr,-Glu-Leu and extending to Phew8. Analysis of CB IV provided 20 residues from Ala,7-Ala-Gly to Vale06; Seri03 was identified by conventional analysis (see below).

Alignment of CNBr Fragments. Since the dipeptide CB V

was blocked, it was placed at the amino terminus of the molecule. Tryptic peptides Tp I and Tp III were also blocked; an amino-terminal acetyl group was identified on Tp I. The

)9



1310 Biochemistry: Enfield et al.

LyS Lys * Met Lyes Ph* Lys Met Lye Lys

tI ll l 11' 111)1

.1 1 2 12 \ 18 27 3236 54

13 26

SOURCES OF FRAGMENTS

I Cleavage at methionines with CNBr CB x CB II

'/////////////////////////////A CBm

E/// //////// , I SUBDIGESTS: CHT

m c-2

IT Cleavage at orginine after succinylation

III Tryptic digest

TpI TpI-3 Tp I Tp M

I 1l'. /////1 Tp I Tp

I I 1 "-- TpV

* Detail of peptide sequences and alignment Residue 14-34

15 20 Ala lie Gly Ala Phe Ala Ala Ala Glu Ser P

ceBm TpX-3

Tpir

Residues 58-78

60 65 Ile Glu Glu Glu Glu Leu Gly Phe Ile Leu L

CBU

Tp I3?

Residues 100-109

100 105 109 Asp Glu Phe Ser Thr Leu Val Ser Glu Ser

CB 33?

/fc-4

]C-2

FIG. 1. Summary of the determination of the amino-acid sequen where specific cleavage points are indicated. Only peptides discusse cated by asterisks are detailed in the lower part of the diagram. Re or --, automated degradation; ... or -, manual degradation; - -,

composition of Tp I corresponds to the sum of CB V and the amino-terminal 11 residues of CB III. CB IV contained no homoserine and was thus placed at the carboxyl terminus of the molecule. CB II, therefore, must be located between CB III and CB IV.

Since CB II contains the single arginyl residue, an arginine- specific tryptic cleavage was performed on the intact protein. Sequenator analysis of fragment R I so obtained (Fig. 1) provided the sequence beginning with Asp76-Leu-Ser and extending through Mets8-Ala-Ala-Gly, thus establishing the overlap between CB II and CB IV.

Total Sequence. Sequence analysis of selected tryptic and chymotryptic peptides provided the sequence of the remaining residues of the parvalbumin molecule, as summarized in Fig. 1. The amino-acid compositions of the chymotryptic peptides used are given in Table 3. The sequence of the region of the molecule between Pheig and Met32-Val-Gly was determined by analysis of peptides Tp X-3, Tp XV, III C-2, and Tp IX and confirmed the alignment of fragments CB III and

Proc. Nat. Acad. Sci. USA 72 (1975)

* * Lye Arg Ly Me Ly s M y

I I A' 'i : I I Id 68 75 / 83 86 96 109

1

/////////////////// 1

CHT CHT

aC-19 cC-4 E1 E

ci C-4 IV C-2

[El

RI

Tp .-5

Tp Il

25 30 he Asp His Lys Lys Phe Phe Gin Met Val Gly

Tp IX

mc-2 CBIa

70 75 ys Gly Phe Ser Pro Asp Ala Arg Asp Leu Ser

;-19 RI

Tp33

ce of rabbit parvalbumin. The top bar represents the intact protein, d in the text are shown. Regions enclosed by parentheses and indi- sidue numbers correspond to the sequence in Fig. 3. Crosshatching unidentified residues.

CB II. Peptides Tp V and Tp XV resulted from cleavage of the Phe,8-Ala peptide bond. The high yields of these peptides relative to that of Tp X-3, the expected tryptic peptide, suggests that the hydrolysis of this bond may represent an unusual tryptic cleavage.

Residues 55-75 were determined by analysis of peptides Tp IV, II C-19, Tp XI, and II C-ll. Sequenator analysis of

Tp IV yielded the four glutamyl residues at positions 59-62. Ser55 was identified only at very low levels in three separate sequenator analyses. However, the solution of the PhS- hydantoin-amino acid after the conversion step was deep pink, which is indicative of the presence of PhS-hydantoin- serine (26). The seryl residue was confirmed by subtractive Edman degradation of Tp IV. Furthermore, complete digestion of Tp IV by aminopeptidase M (Rohm and Haas) yielded unmodified serine in 77% yield and confirmed the amino-acid composition. Manual degradation of the arginine-containing chymotryptic peptide II C-ll confirmed that fragment R I had resulted from cleavage at the arginyl residue.

The sequence of the carboxyl terminus of the parvalbumin




I .2

1.0

Ai _ 0.8

,-o

I I 0.6-

5m 0.4 0

0.2 2

20 40 6o0 80 100 120 140

FRACTION NUMBER

FIG. 2. Gel filtration of the CNBr digest of rabbit parvalbumin. The lyophilized digest (60 mg) was applied to a 2.5 X 115 cm column of Sephadex G-50 "superfine" and developed at 20 ml/hr with 9% formic acid. Fractions were collected every 15 min and monitored by ninhydrin analysis of 50-ul aliquots after alkaline hydrolysis. Fractions were pooled as indicated and peptides recovered by lyophilization.

qt molecule was completed by degradation of chymotryptic peptides IV C-4 and IV C-2 derived from the carboxyl- re terminal CNBr fragment CB IV. Residues 97-109 thus corre- P

spond in composition to the carboxyl-terminal tryptic peptide Tp II.

DISCUSSION

The amino-acid sequence of rabbit parvalbumin given in Fig. 3 agrees closely with the composition reported by Lehky et al. p( (17). The compositions of fragments CB 'II through CB V in correspond to that calculated from the sequence. The high ir values of alanine and homoserine in pooled fraction CB III C (Table 1) are due to the presence in this fraction of fragment h? CB V-III, which resisted cleavage of the Met2-Thr bond. re

TABLE 2. Amino-acid compositions of t

Amino acid TpI TplII TplII TpIV TpV TpIX TpX-2 TpX-3

Lys 1.0(1) 2.1(2) 1.2(1) 1.0(1) 1.0(1) 1.0(1) Hlis 1.0(1) Arg Asp 2.0(2) 1. 1(1) 2.0(2) 1.1(1) 1.0(1) Thr 1.0(1) 0.9(1) 0.9(1) Ser 2.7(3) 0.8(1) 0.9(1) 1.0(1) Glt 2.2(2) 2.0(2) 2.0(2) 3.9(4) 1.0(1) 1.0(1) Pro

Gly 1.0(1) 2.0(2) 1.0(1) 1.1(1) 1.0(1) Ala 2.0(2) 1.0(1) 1.8(2) 2.1(2) 4.7(5) Val 0.8(1) 1.0(1) 1.1(1) Met 0.9(1) 0.9(1) 0.9(1) lie 1.0(1) 1.0(1) 0.9(1) 1.8(2) 1.0(1) 1.0(1) Let 1.9(2) 1.0(1) 2.0(2) 2.0(2) 1.0(1) 1.0(1) Phe 1.0(1) 2.0(2) 1.0(1) 1.8(2) 1.9(2)

Total 12 13 13 14 5 8 5 14 % Yield 52 100 34 36 37 68 44 19 Residue

no. 1-12 97-109 1-13 55-68 14-18 29-36 76-80 14-27

* Residues per molecule. Integral values determined by sequence anal to the sequence given in Fig. 3.

t From a digest of one preparation of parvalbumin, both TpX-5 and at its amino terminus (position 84), were isolated at a molar ratio of 2:1

Sequence of Rabbit Parvalbumin 1311

TABLE 1. Amino-acid compositions of CNBr fragments from rabbit parvalbumin*

Total Rabbit mino in se- parval- ,cid CB II CB III CB IV CB V quence bumint

Lys 9.7(10) 4.0(4) 2.0(2) 16 16.1 lis 0.9(1) 0.9(1) 2 2.0 krg 1.0(1) 1 1 .2 ksp 4.8(5) 3.0(3) 4.0(4) 12 12.2 rhr 2.9(3) 1.0(1) 1.0(1) 5 5.2 mer 3.6(4) 1.0(1) 2.7(3) 8 8.0 Ise 0.9(1) 1.5(1)t 0.9(1) 3? 2.8? }lu 5.8(6) 3.8(4) 2.0(2) 12 12.7 Ero 1.0(1) 1 0.8 Sly 4.0(4) 1.1(1) 3.9(4) 9 9.4 kla 1.3(1) 6.8(6)t 3.2(3) 1.0(1) 11 11.3 Val 4.2(4) 1.1(1) 5 5.5 le 2.9(3) 2.0(2) 1.0(1) 6 6.0 Leu 5.7(6) 1.9(2) 1.0(1) 9 9.0 ?he 3.9(4) 4.0(4) 1.0(1) 9 9.1

Fotal 54 30 23 2 109

* Residues per molecule. Integral values determined by se- ience analysis are given in parentheses. t Taken from ref. 9. The composition was normalized to two sidues of histidine. The protein contains no cysteine, trypto- ian, or tyrosine. t CB III appears to contain equal quantities of two frag- ents; one includes residues 3-32, the other the blocked sequence -32 (see text). No methionine was observed. ? Methionine values.

Digestion of parvalbumin with trypsin followed the ex- ected enzymatic specificity except for the unusual cleavage high yield of the Phels-Ala bond. If the position of this bond the tertiary structure is the same as that of the analogous

ysi8-Ala bond in carp parvalbumin, the bond is internal and rdrolysis could not occur without denaturation or prior lease of the tryptic peptide, Tp X-3.

ryptic peptides from rabbit parvalbumin*

fpX-5t TpX-6 TpXI TpXV TpXVII TpXVIII TpXIX TpXX TpXXII

2.0(2) 2.0(2) 1.2(1) 1.1(1) 1.1(1) 1.0(1) 1.1(1) 2.1(2) 1.0(1) 1.0(1) 0.9(1)

1.0(1) 3.0(3) 1. i(1) 1.1(1) 1.1(1) 1.0(1) 1.0(1) 2.1(2) 1.0 ( (1) 0.9(1) 1.0(1)

1.0 (1) 0.9(1) 0.9(1) 1.0(1) 1 .0(1) 1.0(1)

0.9(1) 2.8(3) 1.1(1) 2.0(2) 1.0(1) 2.8(3)

0.9(1) 1.0(1) 1.0(1) ).9(1)

0.8(1) 0.9(1) 1.0(1) 1.0(1) 1.0(1)

1.0(1) 1.0(1) 1.1(1) 1.1(1)

13 7 7 9 3 2 1 7 9 29 36 86 46 48 22 164 7 43

84-96 39-45 69-75 19-27 81-83 53-54 13, 28 46-52 46-54 37, 38

ysis are given in parentheses. The residue numbers correspond

I similar peptide, containing phenylalanine instead of threonine



1312 Biochemistry: Enfield et al.

1 . Carp Phe Ala Gly Val Hake Phe Ala Gly Ile Pike Lys Asp Leu Rabbit Ac-ALA MET-THR-GLU - LEU-

16 25 G u Cys Lys Asp Ala Cys Lys Glu Gly Lys Asp Val Lys Glu Gly Asn GLY-ALA-PHE-ALA-ALA-ALA-GLU-SER-PHE- ASP-

36 45 Thr Ser Ala Asp

Gly Ala Ala Ile Ala Met Ala Asn

LYS-LYS - LYS -SER -THR -GLU- ASP-VAL- LYS- LYS -

56 65 Asp Lys Leu

Asp Val Asp Lys Leu Lys

GLY - - IILE - G GLU - GLU - GLU LEU-GLY - PHE -

76 85 Ala Thr Asp Gly Phe Ala Thr Asp Ala Ala Phe

Thr Asp Ala Ala Phe ASP -LEU -SER -VAL -LYS -GLU -THR-LYS -THR-LEU-

96 105 Val Thr Ala Val Glu Ala Ala Met Ile Glu

LYS-ILE -GLY- ALA -ASP G- PHE-SER -THR-LEU -

FIG. 3. Amino-acid sequence of rabbit skeletal muscle parvalbun and pike III (13) parvalbumins. The sequence of the rabbit parvalbun that differ from those of the rabbit are shown. Residues of the rabb of carp III (15) are enclosed; residues corresponding to those formin, bumins from carp and hake contain 108 residues, whereas those from

The sequence of rabbit parvalbumin is compared in Fig. 3 to those of carp, hake, and pike to demonstrate the homologous relationship among these proteins. All of the residues serving as calcium ligands in carp parvalbumin (15) occupy identical positions in the rabbit protein. Residues neighboring these ligand sites are also highly conserved. Likewise, Arg97 and Glus8, which form the internal salt bridge and play an

TABLE 3. Amino-acid compositions of chymotryptic peptides from CNBr fragments of rabbit parvalbumin*

Amino acid IIC-11 IIC-19 IIIC-2 IVC-2 IVC-4

Lys 1.1(1) 2.1(2) His 1.0(1) Arg 1.0(1) Asp 2.0(2) 1.0(1) Thr 1.0(1) Ser 1.0(1) 1.8(2) 1.0(1) Glu 1.0(1) Pro 1.0(1) Gly 1.0(1) Ala 1.1(1) Val 1.0(1) Leu 1.0(1) 1.0(1) Phe 1.0(1) 1.1(1) Total 7 3 5 4 3 % Yield 28 30 10 40 34 Residue

no. 71-77 68-70 25-29 106-109 103-105

* Residues per molecule. Integral values determined by sequence analysis are given in parentheses. The residue numbers correspond to the sequence given in Fig. 3.


10 15 Asp Ala Ala Ala Leu

Ala Asp Ala Thr Alo Leu Lys Ala Asp Asp Ile Lys Leu Leu LEU-ASN-ALA-GLU-ASP-ILE -LYS -LYS -ALA-ILE-

35 Ala Ala Lys

Gly Glu Thr Lys Ile Ala Ala Lys

HI S - LYS - LYS- PHE- PHE- GLN - MET-VAL- GLY- LEU -

55 Ala Ala Ile Gin

Gly Ile Gin Lys Ala Ile Ala Ala

VAL-PHE-HIS-ILE- LEU _SP-LYS -A LYS-ER

75 Phe Gin Asn Lys Ala Phe Gin Asn Ala Gly Val Ser Ala Ala Gly ILE-LEU-LYS-GLY-PHE -SER - PRO-ASP-ALA-ARG

95 Leu Lys Ser Leu Lys Ser Leu Lys Ala

MET-ALA-ALA-GLY -ASP- LYS -ASP-GLY - A- GLY-

109 Lys Ala Lys Gly His Ala

VAL- SER -GLU -SER

in. The sequence is compared to those of carp III (10), hake (12), in is given in capital letters; of the other parvalbumins, only residues it parvalbumin corresponding to those involved in calcium binding

the hydrophobic core of carp III (15) are underlined. The parval- pike and rabbit have an additional residue at the carboxyl terminus.

essential role in determining the tertiary structure of the molecule, are conserved, as are all of the residues whose sidechains are involved in the formation of hydrogen bonds in the region of this salt bridge (15), except for the substitution of glutamic acid for aspartic acid at position 22. Finally, residues composing the hydrophobic core of carp parvalbumin (15) are either conserved in the rabbit protein or replaced by other hydrophobic residues.

Comparison of the amino-acid sequence of rabbit parvalbumin with that of rabbit TN-C (14) confirms (a) that the two proteins are structurally similar, supporting earlier sug- gestions (14, 27, 28) that they might have evolved from a common ancestor, and (b) that the parvalbumins are not formed simply by limited proteolysis of the TN-C molecule (27, 29). Alignment of the sequences of rabbit parvalbumin and TN-C reveals a degree of identity similar to that found in the comparison of the parvalbumins of carp, hake, and pike with rabbit TN-C (14). Again, maximum identities between the two proteins are observed in the regions implicated in the binding of calcium, especially the carboxyl-terminal seg- ments.

The fact that parvalbumins are not restricted to aquatic lower vertebrates but have been conserved through close to 500 million years to evolution suggests that they must serve some essential physiological function, possibly related to the contractile process.

Analysis of the sequence of rabbit parvalbumin is being carried out independently in the laboratory of Dr. J.-F. Pechere, Department of Macromolecular Biochemistry, CNRS, Montpellier, France.

We acknowledge the excellent technical assistance of Brita Moody, Dr. Albert Boosman, Richard Granberg, and Richard Olsgaard. We thank Dr. P. Lehky for providing the rabbit




parvalbumin used in this investigation. We also thank Drs. K. A. Walsh and K. Titani for valuable discussion. This work was supported in part by the National Institutes of Health (GM 15731 and AM 07902), the National Science Foundation (GB 20482), the Muscular Dystrophy Association of America, and the American Cancer Society (BC91P). D.L.E. is an Associate Investigator of the Howard Hughes Medical Institute.

1. Deuticke, H. J. (1934) Hoppe-Seyler's Z. Physiol. Chem. 224, 216-228.

2. Focant, B. & Pechere, J.-F. (1965) Arch. Int. Physiol. Biochim. 73, 334-354.

3. Hamoir, G. & Konosu, S. (1965) Biochem. J. 96, 85-97. 4. Konosu, S., Hamoir, G. & Pechere, J.-F. (1965) Biochem.

J. 96, 98-112. 5. Pechere, J.-F., Demaille, J. & Capony, J.-P. (1971) Bio-

chim. Biophys. Acta 236, 391-408. 6. Benzonana, G., Capony, J.-P. & Pechere, J.-F. (1972)

Biochim. Biophys. Acta 278, 110-116. 7. Gerday, C. & Teuwis, J.-C, (1972) Biochim. Biophys. Acta

271, 320-331. 8. Pechere, J.-F., Capony, J.-P. & Demaille, J. (1973) Syst.

Zool. 22, 533-548. 9. Bushana Rao, K. S. P. & Gerday, C. (1973) Comp. Biochem.

Physiol. B 44, 1113-1125. 10. Coffee, C. J. & Bradshaw, R. A. (1973) J. Biol. Chem. 248,

3305-3312. 11. Pechere, J.-F.,, Capony, J.-P. & Ryden, L. (1971) Eur. J.

Biochem. 23, 421-428. 12. Capony, J.-P., Ryden, L., Demaille, J. & Pechere, J.-F.

(1973) Eur. J. Biochem. 32, 97-108.

Sequence of Rabbit Parvalbumin 1313

13. Frankenne, F., Joassin, L. & Gerday, C. (1973) FEBS Lett. 35, 145-147.

14. Collins, J. H. (1974) Biochem. Biophys. Res. Commun. 58, 301-308.

15. Kretsinger, R. H. & Nockolds, C. E. (1973) J. Biol. Chem. 248, 3313-3326.

16. Hendrickson, W. A. & Karle, J. (1973) J. Biol. Chem. 248, 3327-3334.

17. Lehky, P., Blum, H. E., Stein, E. A. & Fischer, E. H. (1974) J. Biol. Chem. 249, 4332-4334.

18. Pechbre, J.-F. (1974) C.R. Acad. Sci. 278, 2577-2579. 19. Titani, K., Hermodson, M. A., Fujikawa, R., Ericsson,

L. H., Walsh, K. A., Neurath, H. & Davie, E. W. (1972) Biochemistry 11, 4899-4903.

20. Hermodson, M. A., Ericsson, L. H., Neurath, H. & Walsh, K. A. (1973) Biochemistry 12, 3146-3153.

21. Hermodson, M. A., Ericsson, L. U., Titani, K., Neurath, H. & Walsh, K. A. (1972) Biochemistry 11, 4493-4502.

22. Shearer, W. T., Bradshaw, R. A., Gurd, F. R. N. & Peters, T., Jr. (1967) J. Biol. Chem. 242, 5451-5459.

23. Schmer, G. & Kreil, G. (1969) Anal. Biochem. 29, 186- 192

24. Ambler, R. P. (1965) Biochem. J. 96, 32P. 25. Jackson, R. L. & Hirs, C. H. W. (1970) J. Biol. Chem.

245, 624-636. 26. Ingram, V. M. (1953) J. Chem. Soc., 3717-3718. 27. Malencik, D. A., Heizmann, C. W. & Fischer, E. H. (1975)

Biochemistry, in press. 28. Demaille, J., Dutruge, E., Eisenberg, E., Capony, J.-P. &

Pechere, J.-F. (1974) FEBS Lett. 42, 173-178. 29. Heizmann, C. W., Malencik, D. A. & Fischer, E. H. (1974)

Biochem. Biophys. Res. Commun. 57, 162-168.



amino-acid sequence of parvalbumin from rabbit skeletal muscle

Documents