Large conductance Ca²⁺-activated potassium (BK) channels are composed of pore forming subunits and auxiliary subunits. The subunits are widely expressed in many cell types, while the subunits are more tissue specific and influence diverse aspects of channel function. In the current study we identified the presence of the smooth muscle specific 1 subunit in murine colonic tissue using Western blotting. The native 1 subunits migrated in SDS-PAGE as two molecular weight bands. Enzymatic removal of N-linked glycosylations from the 1 subunit resulted in a single band that migrated lower than the native 1 subunit bands, suggesting that the native 1 subunit exists in either a core glycosylated or highly glycosylated form. We investigated the functional consequence of deglycosylating the 1 subunit during inside-out single channel recordings. During inside-out single channel recordings, with N-Glycosidase F in the pipette solution, the open probability (P_o) and mean open time of BK channels increased in a time dependent manner. Deglycosylation of the BK channel did not affect the conductance, but shifted the steady-state voltage of activation toward more positive potentials without affecting slope when Ca²⁺ concentration were below 1 µM. Treatment of myocytes lacking the 1 subunits of the BK channel with N-Glycosidase F had no effect. These data suggest that glycosylations on the 1 subunit in smooth muscle cells can modify the biophysical properties of BK channels.