Porcine pancreatic lipase. Completion of the primary structure

De, Josiane; Boudouard, Maryse; Bonicel, Jacques; Guidoni, A.; Desnuelle, P.; Rovery, M.

doi:10.1016/0005-2795(81)90126-4

Cited by 158 publications

(41 citation statements)

References 24 publications

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“…Although it is lower in the other units (data not shown), this suggests that the building block of Tg may have originated from the duplication of a smaller unit. A much weaker internal homology has been described in the 3' region of the rat Tg gene [34]. As no relationship could be observed between these repeats and the 5' repetitive unit described in the present study, it appears that the aminoterminal and the carboxyl-terminal portions of Tg evolved from separate building blocks.…”

Section: Dcontrasting

confidence: 78%

Presence of hormonogenic and repetitive domains in the first 930 amino acids of bovine thyroglobulin as deduced from the cDNA sequence

Mercken

Simons

Martynoff

et al. 1985

European Journal of Biochemistry

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The sequence of the first 2831 nucleotides of bovine thyroglobulin mRNA has been determined from the analysis of a cDNA clone. Following a 41‐nucleotide 5′ untranslated sequence, a single open‐reading frame encoding 930 amino acids was observed. This corresponds to the aminoterminal third of thyroglobulin, preceded by a putative signal peptide of 19 amino acids. The protein sequence was found to be essentially made of the sevenfold repetition of a 60‐amino‐acid‐long building unit, interrupted at fixed positions by unrelated segments of variable length. The presence of an internal homology within the repetitive unit itself suggests that the 5′ region of the thyroglobulin gene has evolved from the initial duplication of a relatively short sequence, followed by the serial duplication of the resulting unit. The tyrosine residue at position five has been assigned an important hormonogenic function [Mercken, L., Simons, M.‐J. and Vassart, G. (1982) FEBS Lett. 149, 285–287]. This residue is flanked by sequence elements related to the repeated unit, suggesting that the hormonogenic domain evolved also from the basic ancestor sequence.

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Section: Dcontrasting

confidence: 78%

Presence of hormonogenic and repetitive domains in the first 930 amino acids of bovine thyroglobulin as deduced from the cDNA sequence

Mercken

Simons

Martynoff

et al. 1985

European Journal of Biochemistry

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“…This research is the first ever achieved in a molecule of lipase. It was performed by using mixtures of isolipases since no allotypic replacement could be detected in their amino acid sequence [2,12]. The total data are illustrated in Fig.…”

Section: Discussionmentioning

confidence: 99%

“…In a first attempt to explain this typical heterogenous biocatalysis process [l] at the molecular level, the porcine enzyme was fully sequenced and found to be formed by a single chain of 449 amino acids including 14 half-cystines [2].…”

mentioning

confidence: 99%

Porcine Pancreatic Lipase

Benkouka

Guidoni

Josiane

et al. 1982

European Journal of Biochemistry

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Following complete sequence analysis of the 449 amino acids in porcine pancreatic lipase [J. De Caro et al. (1981) Biochim. Biophys. Acta, 671, 129-1381, the position of the six disulfide bridges and of the two free thiols of the protein was investigated using a variety of techniques. Three bridges (Cys-4 -Cys-10, Cys-237 -Cys-261 and Cys-433 -Cys-449) were easily identified in the peptic digest of lipase at pH 2.0. In the latter digest, two other bridges (Cys-285 -Cys-296 and Cys-299-,Cys-304) were also identified by means of the cystine peptide constituted by two peptide segments: Ala281-Gly-Phe~Pro-Cys-Asp-SerZ87 and Th?92-Ala-Asn-Lys-Cys-Phe-Pro-Cys-Pro-Ser-GluGly-Cy~-Pro-Gln-Met~~'. A disulfide bridge, formed by Cys-285 and one of the three half-cystines of the other segment, connected the two peptide moieties. A second disulfide bridge linked the two remaining half-cystines. It was not possible to split any peptide bond between Cys-296 and Cys-304 with proteolytic enzymes. The determination of the pairing of the four half-cystines was resolved as follows. Bond Lys-295 -Cys-296 was cleaved with trypsin. A single cycle of Edman degradation was then performed on the peptide compound, thus freeing, in particular, Cys-296 of the peptide bond (296-297). Cys-296 only retained its S-S connection with the half-cystine partner. At the completion of the above operations, the two peptide segments (282 -287) and (297 -307) could be separated. Consequently it was concluded that Cys-285 is linked to Cys-296. Therefore Cys-299 and Cys-304 are paired.The most reactive SH, group of the enzyme was localized on Cys-181 by condensation of the native protein with radioactive N-ethylmaleimide. The alkylation of the SHll group required previous denaturation of the molecule at alkaline pH. ,The results obtained for the characterization of the SHll group suggested the existence of two isomeric forms, the SHll being either on Cys-101 or Cys-103 and the bridge alternately between Cys-90-Cys-103 or Cys-90-Cys-101.It is not yet known whether these two forms pre-exist in native lipase or result from an exchange reaction. The bridge Cys-90-Cys-101 was characterized in a thermolytic digest of a cyanogen bromide fragment (CN 11) of the protein. However, another bridge involving Cys-181 was also found in the digest. This bridge is considered as being an artefact. It is possible that considering the treatments undergone by the large peptide (CN 11), the SH groups of lipase were oxidized and transformed in an S-S bridge.The disulfide bridges of lipase form relatively small loops along the main chain. This arrangement is consistent with a high flexibility of the molecule. 580-5921, the SHI group is not essential for lipase activity. The role of the SHrr group should be more precisely investigated.In the duodenum, pancreatic lipase (triacylglycerol acyl hydrolase) very rapidly hydrolyzes natural water-insoluble triacylglycerols into monoacyl glycerols and free fatty acids which are later taken up by the intestinal mucosa. In a first attempt to ...

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“…esterification etc) provided that water should be continually removed (Salleh et al, 2006). It is generally accepted that lipases carry out catalysis via a catalytic triad consisting of the residues S 162 , H 263 and D 176 (porcine pancreas lipase numbering) (De Caro et al, 1981), though there are cases where an E has replaced the D residue. Although, there is little homology among the known sequences of lipases, however, there are evidences indicating the convergent nature of the catalytic motifs of serine proteases and lipases (Ollis et al, 1992).…”

Section: Catalytic Motifs and Sequences Of Two Three Etc Subsitesmentioning

confidence: 99%