Evolution

Sequence Homology

Materials and Methods

The following query protein sequence of N-acetylneuraminic Acid was obtained from GenBank:

N-acetylneuraminic Acid

mgsdkihhhh hhmglsrvra vffdldntli dtagasrrgm levikllqsk yhykeeaeii cdkvqvklsk ecfhpystci tdvrtshwee aiqetkggad nrklaeecyf lwkstrlqhm iladdvkaml telrkevrll lltngdrqtq rekieacacq syfdaivigg eqkeekpaps ifyhccdllg vqpgdcvmvg dtletdiqgg lnaglkatvw inksgrvplt sspmphymvs svlelpallq sidckvsmsv

Fig. 1: Amino Acid Sequence (260 aa) of N-acetylneuraminic Acid (2gfh) protein from Mus musculus (House Mouse)

A total of 500 proteins were yielded. Only a 38 proteins were selected for comparison as these had shown higher homology to the query sequence, in contrast with the remainder of the search results.

These proteins were chosen according to their bit scores and E-values. Two more outlier partial sequences contributing to poor overall alignment (huge deletion gaps) were subsequently removed. The remaining 36 sequences were used for the generation of the phylogenetic tree (and bootstrapped tree as well).

The bacteria sequence matches used for multiple sequence analysis and phylogenetic tree mapping were representative of the other baterial sequences not selected. These selected sequences had the highest bit scores and E-values.

Multiple Sequence Alignments (msa)

Materials and Methods

The following sequence alignment was obtained (Fig. 2)

From the alignments, gi|10888xy and gi|10888yz are representative of gi|108881764 and gi|108881765 respectively. Both these hypothetical proteins belong to the mosquito Aedes aegypti.

The identifier numbers for these two proteins were initially changed to an alpha-numeric one, due to the inability of Phylip to generate a tree from the original identifiers. This was due to the fact that the programme only took the first five numeric digits (10888), thereby resulting in a programme error prompt which listed both proteins as duplicates (from the identifier numbers). Both these identifiers were subsequently renamed for the final phylogenetic tree.

Figure 1. Multiple Sequence Alignments of 2gfh amino acid sequences with that of other similar protein sequences in various organisms (A) Glycolysis pathway with substrates that are hydrolyze by HADs: glucose 6-phosphate, fructose 6-phosphate and dihydroxyacetone phosphate. (B) Pentose phostphate pathway with substrates that are hydrolyze by HADs: glucose-6-phosphate, fructose-6-phosphate, dihydroxyacetone phosphate, glyceraldehyde-3-phosphate, gluconate 6-phosphate and erythrose-4-phosphate.

Phylogenetic Map (Tree)

Table of functions

Catalytic activity	N-acylneuraminate 9-phosphate + H2O = N-acylneuraminate + phosphate
Cofactor	Magnesium (By similarity)
Enzyme regulation	Inhibited by vanadate and calcium (By similarity)
Pathway	Carbohydrate metabolism; aminosugar metabolism
Similarity	Belongs to the haloacid dehalogenase-like hydrolase superfamily. NANP family

Separate Enzymes Catalyze the Final Two Steps of Coenzyme A Biosynthesis in Brevibacterium ammoniagenes: Purification of Pantetheine Phosphate Adenylyltransferase