Newcastle wrapped up the biggest deal of the day, signing England striker Michael Owen from Real Madrid for £17m.
They also re-signed Nolberto Solano and let Jermaine Jenas leave for £7m to Spurs, who also signed striker Grzegorz Rasiak from Championship side Derby.
Middlesbrough agreed to sign midfielder Fabio Rochemback and Portsmouth signed Dario Silva in other last-minute deals.
Celtic and Rangers were also involved in late moves - respectively signing Du Wei and Filippo Maniero.
Transfer news as it happened (All times BST):
0000: The transfer window slams shut until December 31 - with Football League clubs experiencing it for the first time.
2351: Spurs swoop to sign Polish striker Grzegorz Rasiak from Championship outfit Derby County.
2334: Charlton sign former Arsenal, Coventry and Perugia striker Jay Bothroyd.
2330: Nolberto Solano has passed a medical and returns to Newcastle in a £1.5m move from Aston Villa, who will take James Milner on loan for the season.
2300: Portsmouth make a third signing with the capture of Salif Diao on a season-long loan from Liverpool.
2220: Rangers defender Michael Ball joins PSV Eindhoven on two-year contract, with option for a further year.
2210: Celtic sign China international defender Du Wei on a four-year deal.
2140: Nolberto Solano is having a medical at secret location with a view to a move back to Newcastle from Aston Villa. James Milner is expected to go the other way on a season-long loan.
2115: Middlesbrough agree deal to sign Brazilian midfielder Fabio Rochemback from Barcelona.
2100: Uruguayan striker Dario Silva completes a move to Portsmouth from Spanish club Sevilla.
2015: Liverpool's attempts to sign Benfica striker Simao Sabrosa fall through.
1950: Bolton complete the signing of midfielder Fabrice Fernandes, who was released by Southampton, on a free.
1900: Burnley secure the services of defender Duane Courtney on a two-year contract from Telford.
1815 Blackburn sign midfielder David Bentley on loan from Arsenal until end of season.
1800: Leicester City announce a double signing, bringing in Tranmere's Iain Hume on a £500,000 deal rising to £750,000 if the Foxes are promoted - and also signing Fulham's Elvis Hammond in a deal which could net the Cottagers up to £350,000.
1730: Tottenham confirm that Erik Edman has left the club to join French side Rennes.
1700: Window closes for deals between Football League clubs.
1630: Portsmouth confirm the signing of Barcelona's France Under-19 midfielder Frank Songo'o.
1625: Everton announce that they have completed the signing of Dutch international midfielder Andy van der Meyde from Inter Milan for an undisclosed fee.
1545: - Scottish Premier League leaders Hearts have signed Brazilian midfielder Samuel Almeida Camazzola.
1525: Sunderland goalkeeper Mart Poom joins Arsenal on loan.
Going the other way is defender Justin Hoyte, who swaps Highbury for Wearside on a loan deal until the end of the season.
1520: Everton have still to decide whether to spend £2m on injured Dutch winger Andy van der Meyde, and they are also chasing a striker.
1500: Republic of Ireland international forward Connolly joins Wigan from Leicester for £3m.
1330: Tottenham complete the signing of defender Lee Young-Pyo from PSV Eindhoven.
1305: Blackburn have signed Georgian defender Zurab Khizanishvili from Rangers on a season long-loan.
The deal includes an option for Blackburn to buy the player between now and the end of May.
1245: Middlesbrough striker Joseph-Desire Job moves to Saudi Arabian side Al Ittihad on a year's loan.
1215: Simao Sabrosa appears set to join Liverpool from Benfica after the Portuguese Football Federation gave permission for the forward to undergo a medical at Anfield.
PSV Eindhoven defender Lee Young-Pyo has signed for Tottenham, according to his agent.
1200: Owen signs a four-year deal with Newcastle, who fought off Liverpool's interest in the 25-year-old striker.
1045: Middlesbrough defender Michael Reiziger is expected to sign for Dutch side PSV Eindhoven.
1015: Jenas completes his switch from Newcastle to Tottenham.
1000: Tottenham are at the centre of the day's early activity.
The Londoners are set to bring in Jenas for £8m after Newcastle confirmed the clubs had reached agreement over the midfielder.
But defender Edman is on the verge of joining Rennes, with the French team claiming on their website that the deal has been completed.
0900: A busy day is expected as clubs scramble to make acquisitions and offload players before the transfer window shuts.
Among those set to be involved is Aston Villa's Nolberto Solano, who could re-sign for Newcastle.
And Tottenham's Robbie Keane could switch to Everton, while the Londoners are considering bringing in Cristiano Lucarelli from Italian side Livorno.
Celtic are waiting to learn whether China defender Du Wei will complete an £850,000 transfer from Shanghai Shenhua.