β₁-adrenergic receptors (β₁ARs) are the principle mediators of catecholamine action in cardiomyocytes. We previously showed that the β₁AR extracellular N-terminus is a target for post-translational modifications that impact on signaling responses. Specifically, we showed that the β₁AR N-terminus carries O-glycan modifications at Ser³⁷/Ser⁴¹, that O-glycosylation prevents β₁AR N-terminal cleavage, and that N-terminal truncation influences β₁AR signaling to downstream effectors. However, the site(s) and mechanism for β₁AR N-terminal cleavage in cells was not identified. This study shows that β₁ARs are expressed in cardiomyocytes and other cells types as both full-length and N-terminally truncated species and that the truncated β₁AR species is formed as a result of an O-glycan regulated N-terminal cleavage by ADAM17 at R³¹↓L³². We identify Ser⁴¹ as the major O-glycosylation site on the β₁AR N-terminus and show that an O-glycan modification at Ser⁴¹ prevents ADAM17-dependent cleavage of the β₁-AR N-terminus at S⁴¹↓L⁴², a second N-terminal cleavage site adjacent to this O-glycan modification (and it attenuates β₁-AR N-terminal cleavage at R³¹↓L³²). We previously reported that oxidative stress leads to a decrease in β₁AR expression and catecholamine responsiveness in cardiomyocytes. This study shows that redox-inactivation of cardiomyocyte β₁ARs is via a mechanism involving N-terminal truncation at R³¹↓L³² by ADAM17. In keeping with the previous observation that N-terminally truncated β₁ARs constitutively activate an AKT pathway that affords protection against doxorubicin-dependent apoptosis, overexpression of a cleavage resistant β₁AR mutant exacerbates doxorubicin-dependent apoptosis. These studies identify the β₁AR N-terminus as a structural determinant of β₁AR responses that can be targeted for therapeutic advantage.