Ryanodine receptors (RyR) are intracellular channels that are intricately involved in Ca2+ release. These channels large membrane proteins~2.26MDa in size. In this multi-goal project firstly we successfully studied the gating mechanics of the RyR1 in the presence of Mg2+. We used single particle reconstruction and image processing techniques to obtain the 3D structure of the RyR1 with Mg2+. The 3D structure in the presence of Mg2+ and an ATP analog is the closest representation of human physiological conditions. The open and closed state structures of RyR1 are known. However, the physiologically closed state has not been studied before. Understanding this structure will help in the understanding of protein interactions. Our second goal was the validation of this 3D structure under different levels of noise. Validation under different noise levels analyzed the problem of noise bias is present in the field of cryo-EM and single particle reconstruction in select cases.