Use of carboxypeptidase Y for carboxy-terminal sequence determination in proteins

In this paper, we present the amino-terminal sequence of rat tonin, an endopeptidase responsible for the conversion of angiotensinogen, the tetradecapeptide renin substrate, or angiotensin I to angiotensin II. It is shown that isoleucine and proline occupy the amino- and carboxy-terminal residues respectively. The N-terminal sequence analysis permitted the identification of 34 out of the first 40 residue s of the single polypeptide chain composed of 272 amino acids. The se results showed an extensive homology with the sequence of many serine proteases of the trypsin–chymotrypsin family. This information, coupled with the slow inhibition of tonin by diisopropylfluorophosphate, classified this enzyme as a selective endopeptidase of the active serine protease family.

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Automated Carboxy-Terminal Sequence Analysis of Polypeptides Containing C-Terminal Proline

Analytical Biochemistry ◽

10.1006/abio.1995.1091 ◽

1995 ◽

Vol 224 (2) ◽

pp. 588-596 ◽

Cited By ~ 13

Author(s):

J.M. Bailey ◽

O. Tu ◽

G. Issai ◽

A. Ha ◽

J.E. Shively

Keyword(s):

Sequence Analysis ◽

Terminal Sequence ◽

Carboxy Terminal

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STUDIES ON THE USE OF SILYL COMPOUNDS FOR PROTEIN CARBOXY-TERMINAL SEQUENCE ANALYSIS

Techniques in Protein Chemistry ◽

10.1016/b978-0-12-682001-0.50014-2 ◽

1989 ◽

pp. 67-78 ◽

Cited By ~ 5

Author(s):

Chad G. Miller ◽

Cheng-te Kong ◽

John E. Shively

Keyword(s):

Sequence Analysis ◽

Terminal Sequence ◽

Carboxy Terminal

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The DNA sequence of the structural gene of gonococcal protein III and the flanking region containing a repetitive sequence. Homology of protein III with enterobacterial OmpA proteins.

Journal of Experimental Medicine ◽

10.1084/jem.165.2.471 ◽

1987 ◽

Vol 165 (2) ◽

pp. 471-482 ◽

Cited By ~ 28

Author(s):

E C Gotschlich ◽

M Seiff ◽

M S Blake

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Repetitive Sequence ◽

Consensus Sequence ◽

Terminal Sequence ◽

Inverted Orientation ◽

Amino Acid Signal Peptide ◽

Flanking Region ◽

Carboxy Terminal ◽

Amino Acid Signal

The insert of a lambda gt11 clone expressing gonococcal protein III was sequenced. The deduced amino acid sequence showed a coding frame of 236 amino acids with a typical 22-amino-acid signal peptide, followed by the known NH2-terminal sequence of PIII. The mature protein has a molecular weight of 23,298. It was found that PIII had extensive and very striking homology to the carboxy-terminal portion of enterobacterial OmpA proteins. The homology encompasses the OmpA domain that is believed to be located in the periplasmic space. If the disposition of PIII across the OM is analogous, then the surface-exposed domain consists of less than 40 amino acids. These include a potential 15-amino-acid disulfide loop, a feature not found in OmpA proteins. Hybridization studies with the sequenced insert indicated that it contained a repetitive sequence that occurred at least 20 times in the genome. By additional hybridization studies the area containing the repetitive sequence was narrowed to a region of 43 bp. This region contained an exact copy of the consensus sequence of a 26-bp repetitive sequence recently described. An analogous sequence recurs in an inverted orientation 53 bp downstream.

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