Complete amino acid sequence and location of the five disulfide bridges in porcine pancreatic α-amylase

Paséro, L.; Mazzéi-Pierron, Yvette; Abadie, B.; Chicheportiche, Yves; Marchis-Mouren, G.

doi:10.1016/0167-4838(86)90289-x

Cited by 109 publications

(62 citation statements)

References 21 publications

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“…Based on these alignments, the unrooted distance trees were constructed by the neighbour-joining method [20]. The cysteine residues involved in probable disulfide bridges of the third group of a-amylases represented by streptomycetes and animals were extracted from the amino acid sequence alignment of this group by using the known cystines of pig pancreatic a-amylase [22] as the template.…”

Section: Methodsmentioning

confidence: 99%

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Sequence Similarities and Evolutionary Relationships of Microbial, Plant and Animal α‐amylases

Janeček

1994

European Journal of Biochemistry

100

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Amino acid sequence comparison of 37 a-amylases from microbial, plant and animal sources was performed to identify their mutual sequence similarities in addition to the five already described conserved regions. These sequence regions were examined from structure/function and evolutionary perspectives. An unrooted evolutionary tree of a-amylases was constructed on a subset of 55 residues from the alignment of sequence similarities along with conserved regions. The most important new information extracted from the tree was as follows: (a) the close evolutionary relationship of Alteromonas haloplanctis a-amylase (thermolabile enzyme from an antarctic psychrotroph) with the already known group of homologous a-amylases from streptomycetes, Thermomonospora cuwata, insects and mammals, and (b) the remarkable 40.1 % identity between starch-saccharifying Bacillus subtilis a-amylase and the enzyme from the ruminal bacterium Butyrivibrio fibrisolvens, an a-amylase with an unusually large polypeptide chain (943 residues in the mature enzyme). Due to a very high degree of similarity, the whole amino acid sequences of three groups of a-amylases, namely (a) fungi and yeasts, (b) plants, and (c) A. haloplanctis, streptomycetes, T cuwata, insects and mammals, were aligned independently and their unrooted distance trees were calculated using these alignments. Possible rooting of the trees was also discussed. Based on the knowledge of the location of the five disulfide bonds in the structure of pig pancreatic a-amylase, the possible disulfide bridges were established for each of these groups of homologous a-amylases.Although all a-amylases catalyze the hydrolysis of a-l,4- [lS]. For all these examples, sequence analysis remains to be performed.Therefore, the main goal of the present study was to summarize the findings resulting from the analysis of amino acid sequences of available microbial, plant and animal a-amylases. To further elucidate the evolutionary aspects of a-amylases, unrooted distance trees were constructed and discussed. MATERIALS AND METHODSAmino acid sequences of a-amylases (Table 1)

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Section: Methodsmentioning

confidence: 99%

“…the alignment of cysteine residues when compared with these residues of pig pancreatic a-amylase, which is known to contain five disulfide bridges [22]. All of these bridges are present probably only in the a-amylases of mammals.…”

Section: Evolution Of A-amylasesmentioning

confidence: 99%

Sequence Similarities and Evolutionary Relationships of Microbial, Plant and Animal α‐amylases

Janeček

1994

European Journal of Biochemistry

100

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“…porcine (Pasero et al, 1986;Qian et al, 1993), Taka (Toda et al, 1982), and Niger (Takkinen et al, 1983) a-amylases have been aligned so as to maximize the structural homology present. The single-letter code is used to identify amino acids and the residue numbering is based on the sequence of human pancreatic a-amylase.…”

Section: L a H P V G F T R V M S S Y R W A N F V N G E D V N D W I mentioning

confidence: 99%

The structure of human pancreaticα-amylase at 1.8 Å resolution and comparisons with related enzymes

1995

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The structure of human pancreatic a-amylase has been determined to 1.8 A resolution using X-ray diffraction techniques. This enzyme is found to be composed of three structural domains. The largest is Domain A (residues 1-99, 169-404), which forms a central eight-stranded parallel @-barrel, to one end of which are located the active site residues Asp 197, Glu 233, and Asp 300. Also found in this vicinity is a bound chloride ion that forms ligand interactions to Arg 195, Asn 298, and Arg 337. Domain B is the smallest (residues 100-168) and serves to form a calcium binding site against the wall of the @-barrel of Domain A. Protein groups making ligand interactions to this calcium include Asn 100, Arg 158, Asp 167, and His 201. Domain C (residues 405-496) is made up of antiparallel @-structure and is only loosely associated with Domains A and B. It is notable that the N-terminal glutamine residue of human pancreatic a-amylase undergoes a posttranslational modification to form a stable pyrrolidone derivative that may provide protection against other digestive enzymes. Structure-based comparisons of human pancreatic a-amylase with functionally related enzymes serve to emphasize three points. Firstly, despite this approach facilitating primary sequence alignments with respect to the numerous insertions and deletions present, overall there is only -15% sequence homology between the mammalian and fungal a-amylases. Secondly, in contrast, these same studies indicate that significant structural homology is present and of the order of -70%. Thirdly, the positioning of Domain C can vary considerably between a-amylases. In terms of the more closely related porcine enzyme, there are four regions of polypeptide chain (residues 237-250, 304-310, 346-354, and 458-461) with significantly different conformations from those in human pancreatic a-amylase. At least two of these could play a role in observed differential substrate and cleavage pattern specificities between these enzymes. Similarly, amino acid differences between human pancreatic and salivary a-amylases have been localized and a number of these occur in the vicinity of the active site.Keywords: amylase; crystallography; enzymology; glycogen; pancreatic; sequences; starch; structure a-Amylases catalyze the hydrolysis of a-1,4 glucan linkages in starch and are widely distributed in nature, being found in bacteria, plants and animals. In humans, a-amylase is composed of 496 amino acids in a single polypeptide chain, which is encoded on chromosome 1 as part of a multigene family (Gumucio et al., 1988). These genes are regulated so that different isozymes are synthesized in either salivary glands or the pancreas. The salivary and pancreatic a-amylases are highly homologous in terms of primary sequence (Nishide et al., 1986) but d o exhibit somewhat different cleavage patterns (Minamiura, 1988). The functional differences observed undoubtedly arise from the 15 amino Reprint requests to: Gary D. Brayer, Department of Biochemistry and Molecular Biology, University of B...

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“…In mammals, a-amylase is present in both salivary and pancreatic secretions. Amino acid sequences for mammalian a-amylases show a high degree of homology (Pasero et al, 1986;Qian et al, 1993). Two refined threedimensional structures obtained at 2.1 A resolution have been reported for fungal a-amylases: Aspergillus niger acid amylase and Aspergillus oryzae amylase (Swift et al, 1991).…”

mentioning

confidence: 99%

“…Two isoenzymes (PPA I and 11) are known for pig pancreatic a-amylase (Cozzone et al, 1970). They have the same molecular weight but differ slightly in amino acid composition and isoelectric point (Kluh, 1981;Pasero et al, 1986).…”

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confidence: 99%

Carbohydrate binding sites in a pancreatic α‐amylase‐substrate complex, derived from X‐ray structure analysis at 2.1 Å resolution

1995

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The X-ray structure analysis of a crystal of pig pancreatic a-amylase (PPA, EC 3.2.1.1 .) that was soaked with the substrate maltopentaose showed electron density corresponding to two independent carbohydrate recognition sites on the surface of the molecule. Both binding sites are distinct from the active site described in detail in our previous high-resolution study of a complex between PPA and a carbohydrate inhibitor (Qian M, Buisson G , Duke E, Haser H, Payan F, 1994, Biochemistry 33:6284-6294). One of the binding sites previously identified in a 5-A-resolution electron density map, lies at a distance of 20 A from the active site cleft and can accommodate two glucose units. The second affinity site for sugar units is located close to the calcium binding site. The crystal structure of the maltopentaose complex was refined at 2.1 A resolution, to an R-factor of 17.5%, with an RMS deviation in bond distances of 0.007 A. The model includes all 496 residues of the enzyme, 1 calcium ion, 1 chloride ion, 425 water molecules, and 3 bound sugar rings. The binding sites are characterized and described in detail. The present complex structure provides the evidence of an increased stability of the structure upon interaction with the substrate and allows identification of an N-terminal pyrrolidonecarboxylic acid in PPA.

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Complete amino acid sequence and location of the five disulfide bridges in porcine pancreatic α-amylase

Cited by 109 publications

References 21 publications

Sequence Similarities and Evolutionary Relationships of Microbial, Plant and Animal α‐amylases

Sequence Similarities and Evolutionary Relationships of Microbial, Plant and Animal α‐amylases

The structure of human pancreaticα-amylase at 1.8 Å resolution and comparisons with related enzymes

Carbohydrate binding sites in a pancreatic α‐amylase‐substrate complex, derived from X‐ray structure analysis at 2.1 Å resolution

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