5'-Nucleotidases cleave off the phosphate from the 5' end of the sugar moiety. They can be classified into various kinds depending on their substrate preferences and subcellular localization. Membrane-bound 5'-nucleotidases display specificity toward adenosine monophosphates and are involved predominantly in the salvage of preformed nucleotides and in signal transduction cascades involving purinergic receptors. Soluble 5'-nucleotidases are all known to belong to the haloacid dehalogenase superfamily of enzymes, which are two domain proteins characterised by a modified Rossman fold as the core and variable cap or hood. The soluble forms are further subclassified based on the criterion mentioned above. mdN and cdN are mitochondrial and cytosolic 5'-3'-pyrimidine nucleotidases. cN-I is a cytosolic nucleotidase(cN) characterized by its affinity toward AMP as its substrate. cN-II is identified by its affinity toward either IMP or GMP or both. cN-III is a pyrimidine 5'-nucleotidase. 5'-Nucleotidases are involved in varied functions like cell–cell communication, nucleic acid repair, purine salvage pathway for the synthesis of nucleotides, signal transduction, membrane transport, etc.