The PINCH-1-ILK--parvin (PIP) complex plays important roles in regulation of glomerular cell behavior, including podocyte shape change, apoptosis and mesangial fibronectin matrix deposition. In this study, we show that PINCH-2, a protein that is structurally related to PINCH-1 but encoded by a different gene, is co-expressed with PINCH-1 in podocytes. Treatment of podocytes with TGF-1 elevated the level of PINCH-2, resulting in increased association of PINCH-2 with ILK and -parvin and concomitantly displacement of PINCH-1 from the PIP complex. To gain insights into the functional consequences of elevated PINCH-2 expression, we overexpressed PINCH-2 in podocytes by infection with an adenovirus encoding PINCH-2. Overexpression of PINCH-2 resulted in displacement of PINCH-1 from the PIP complex and compromised podocyte spreading. The PINCH-2-mediated displacement of PINCH-1, however, did not prompt apoptosis. Interestingly, the effect of PINCH-2 on podocyte spreading depends on differentiation status, as overexpression of PINCH-2 in podocytes that were not fully differentiated did not alter cell spreading. Finally, we show that overexpression of PINCH-2 in mesangial cells resulted in displacement of PINCH-1 from the PIP complex but impaired neither mesangial cell spreading nor fibronectin matrix deposition. These studies suggest that PINCH-2 can substitute for PINCH-1 in at least certain processes in glomerular cells (e.g., podocyte survival signaling and mesangial fibronectin matrix deposition), albeit an aberrantly high level of PINCH-2 may contribute to TGF-1-induced alteration in podocyte shape modulation.