SGD applied the ND (No biological Data available) evidence code to AMD2 for all three GO aspects (GO:0003674, GO:0005575, GO:0008150), formally recording that no experimental molecular-function, cellular-component, or biological-process data existed for this gene.

The combined automated IEA pipeline assigns the generic molecular function amidase activity (GO:0004040) to AMD2 without any gene-specific experiment. GO_REF:0000120 is a generic description of the combined-IEA methods and says nothing about AMD2 specifically; the most likely inference route, reconstructed by this review rather than stated by the reference, is the amidase-signature domain together with the EC 3.5.1.4 mapping.

YDR242W/AMD2 was originally reported only as a PUTATIVE amidase gene, identified from genomic sequence similarity to the amidase family; no enzymatic assay or in-vivo characterization accompanied the report.

AMD2 is named only as a "Probable amidase" and is assigned the generic amidase EC number 3.5.1.4; the protein existence level is 3 (inferred from homology), so no protein-level experimental evidence exists.

The annotated catalytic activity is the generic parent reaction of the amidase family, with an unspecified monocarboxylic acid amide as substrate (Rhea:RHEA:12020).

UniProt annotates the amidase-signature domain (Pfam PF01425), the family PROSITE pattern (PS00571), and a Ser-cisSer-Lys catalytic triad at positions 132/209/233 - positions 132 and 209 noted as "Charge relay system" and 233 as the acyl-ester intermediate - with all three ACT_SITE features carrying /evidence="ECO:0000250" (inferred by sequence similarity, not observed). No catalytic residue is annotated as disrupted, so nothing in the sequence argues for pseudoenzyme status, but catalytic competence has not been demonstrated: the entry itself asserts only "Probable amidase" at protein existence level 3.

A functional acetamidase confers growth on acetamide as a sole carbon or nitrogen source by hydrolysing it to acetate and ammonia; growth on acetamide is therefore a direct phenotypic readout for acetamidase activity in a yeast genome.

Untransformed S. cerevisiae S288C - whose genome contains AMD2 - fails to form colonies on acetamide, and acquires the ability to do so only when a heterologous acetamidase (YlAMD1) is supplied, indicating that the native S288C genome, which includes AMD2, does not supply growth-supporting acetamidase activity under these conditions. The readout is whole-genome growth and does not isolate AMD2 itself: the paper does not mention AMD2/YDR242W, and non-expression on this medium, limiting acetamide uptake, or sub-threshold activity would each be consistent with the control failing to grow.