Biochimica et Biophysica Acta, 439 (1976) 51-54

© Elsevier ScientificPublishing Company, Amsterdam-- Printed in The Netherlands BBA 37376 THE PRIMARY STRUCTURE OF THE MYOGLOBIN OF RABBIT ( O R Y C TOLAGUS CUNICULUS)

ROMERO-HERRERA, H. LEHMANN and O. CASTILLO University Department of Clinical Biochemistry, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2QR (U.K.) (Received December 31st, 1975) A. E.

SUMMARY The amino acid sequence of rabbit myoglobin was determined. It contains 153 amino acid residues and differs in three of them from all known mammalian myoglobins: Ala 5, His 34 and Thr 35.

INTRODUCTION We have established the amino acid sequence of the myoglobin of the rabbit (Oryctolagus cuniculus) in an attempt to elucidate the phylogenetic relationships of

this lagomorph. As five antigenic regions of the sperm whale myoglobin molecule have been determined by Atassi [1], we considered it of special interest to study the rabbit myoglobin sequence, because some of the antisera to the sperm whale molecule were prepared in this animal. A detailed account of our phylogenetic and immunological deductions will be published elsewhere. The myoglobin of the rabbit contains 153 amino acid residues, as in all the major fractions of the mammalian myoglobins studied so far, and differs from the sequence of man and sperm whale by 16 and 22 residues, respectively. MATERIALS AND METHODS 500 g of rabbit skeletal muscle were stripped of fat and connective tissue and minced in a blender. After aqueous extraction the myoglobin was purified, as routinely done in our laboratory, by (NH4)zSO4 precipitation [2], DEAE-Sephadex column chromatography [2] and paper electrophoresis [3, 4]. Removal of the haem was achieved by acid acetone [5] and the resulting apomyoglobin was then submitted to CM-cellulose column chromatography [2]. After hydrolysis of the myoglobin in 6 M HC1 for 24, 48 and 72 h at 108 °C, the amino acid composition of the molecule was established in an automatic analyser (Locarte). Quantitation of tryptophan'was performed by hydrolysis with mercaptoethane sulphonic acid [6]. The myoglobin was digested with trypsin [3], pepsin [3] (the insoluble core left after tryptic digestion) and chymotrypsin [3]. The resulting enzymic peptides were separated by fingerprinting (two-dimensional high voltage electrophoresis and

52 1 P-AB~.,'T Mb

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Gly Leu Set Asp ~/a Glu Trp Gln Leu Va/ ~

12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 ASh Val ~ Gly Lla~ Val Glu Ala A~p Ieu Ala Gly HIS Gly

The primary structure of the myoglobin of rabbit (Oryctolagus cuniculus)

Biochimica et Biophysica Acta, 439 (1976) 51-54 © Elsevier ScientificPublishing Company, Amsterdam-- Printed in The Netherlands BBA 37376 THE PRIMARY...
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